Galactopyranosyl-cyclohexyl derivauves as E-selectin antagonists

ABSTRACT

Compounds, compositions, and methods for treatment and/or prevention of at least one disease, disorder, and/or condition by inhibiting binding of an E-selectin to an E-selectin ligand are disclosed. For example, E-selectin antagonists are described and pharmaceutical compositions comprising at least one of the same.

This application is a United States national stage application filed under 35 U.S.C. § 371 of International Patent Application No. PCT/US2018/021977 accorded an international filing date of Mar. 12, 2018; which claims the benefit under 35 U.S.C. § 119(e) to U.S. Provisional Application No. 62/471,860 filed Mar. 15, 2017, all of which are incorporated by reference herein in their entirety.

Compounds, compositions, and methods for treating and/or preventing at least one disease, disorder, and/or condition associated with E-selectin activity including, for example, inflammatory diseases and cancers, are disclosed herein.

When a tissue is infected or damaged, the inflammatory process directs leukocytes and other immune system components to the site of infection or injury. Within this process, leukocytes play an important role in the engulfment and digestion of microorganisms. The recruitment of leukocytes to infected or damaged tissue is critical for mounting an effective immune defense.

Selectins are a group of structurally similar cell surface receptors important for mediating leukocyte binding to endothelial cells. These proteins are type 1 membrane proteins and are composed of an amino terminal lectin domain, an epidermal growth factor (EGF)-like domain, a variable number of complement receptor related repeats, a hydrophobic domain spanning region and a cytoplasmic domain. The binding interactions appear to be mediated by contact of the lectin domain of the selectins and various carbohydrate ligands.

There are three known selectins: E-selectin, P-selectin, and L-selectin. E-selectin is found on the surface of activated endothelial cells, which line the interior wall of capillaries. E-selectin binds to the carbohydrate sialyl-Lewis^(x) (sLe^(x)), which is presented as a glycoprotein or glycolipid on the surface of certain leukocytes (monocytes and neutrophils) and helps these cells adhere to capillary walls in areas where surrounding tissue is infected or damaged; and E-selectin also binds to sialyl-Lewis^(a) (sLe^(a)), which is expressed on many tumor cells. P-selectin is expressed on inflamed endothelium and platelets, and also recognizes sLe^(x) and sLe^(a), but also contains a second site that interacts with sulfated tyrosine. The expression of E-selectin and P-selectin is generally increased when the tissue adjacent to a capillary is infected or damaged. L-selectin is expressed on leukocytes. Selectin-mediated intercellular adhesion is an example of a selectin-mediated function.

Although selectin-mediated cell adhesion is required for fighting infection and destroying foreign material, there are situations in which such cell adhesion is undesirable or excessive, resulting in tissue damage instead of repair. For example, many pathologies (such as autoimmune and inflammatory diseases, shock and reperfusion injuries) involve abnormal adhesion of white blood cells. Such abnormal cell adhesion may also play a role in transplant and graft rejection. In addition, some circulating cancer cells appear to take advantage of the inflammatory mechanism to bind to activated endothelium and metastasize. In such circumstances, modulation of selectin-mediated intercellular adhesion may be desirable.

Accordingly, there is a need in the art for inhibitors of selectin-mediated function, function, e.g., of selectin-dependent cell adhesion, and for the development of methods employing such compounds. The present disclosure may fulfill one or more of these needs and/or may provide other advantages.

Compounds, compositions, and methods for treating and/or preventing (i.e., reducing the likelihood of occurrence or reoccurence) at least one disease, disorder, and/or condition in which inhibiting binding of E-selectin to one or more ligands may play a role are disclosed.

Disclosed are glycomimetic E-selectin antagnoists of Formula (I):

prodrugs of Formula (I), and pharmaceutically acceptable salts of any of the foregoing, wherein R¹, R², R³, R⁴, R⁵, R⁶, L, and M are defined herein.

As used herein, ‘compound of Formula (I)’ includes E-selectin antagonists of Formula (I), pharmaceutically acceptable salts of E-selectin antagonists of Formula (I), prodrugs of E-selectin antagonists of Formula (I), and pharmaceutically acceptable salts of prodrugs of E-selectin antagonists of Formula (I).

In some embodiments, pharmaceutical compositions comprising at least one compound of Formula (I) and optionally at least one additional pharmaceutically acceptable ingredient are presented.

In some embodiments, a method for treatment and/or prevention of at least one disease, disorder, and/or condition where inhibition of E-selectin mediated functions is useful is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments. However, one skilled in the art will understand that the disclosed embodiments may be practiced without these details. In other instances, well-known structures have not been shown or described in detail to avoid unnecessarily obscuring descriptions of the embodiments. These and other embodiments will become apparent upon reference to the following detailed description and attached drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a diagram illustrating the synthesis of building block 3.

FIG. 2 is a diagram illustrating the synthesis of building block 7.

FIG. 3 is a diagram illustrating the synthesis of intermediate 11.

FIG. 4 is a diagram illustrating the synthesis of intermediate 12.

FIG. 5 is a diagram illustrating the synthesis of compound 17.

FIG. 6 is a diagram illustrating the synthesis of intermediate 16.

FIG. 7 is a diagram illustrating the synthesis of compound 24.

FIG. 8 is a diagram illustrating the synthesis of building block 27.

FIG. 9 is a diagram illustrating the synthesis of compound 30.

FIG. 10 is a diagram illustrating the synthesis of compound 33.

FIG. 11 is a diagram illustrating the synthesis of compound 35.

FIG. 12 is a diagram illustrating the synthesis of intermediate 40.

FIG. 13 is a diagram illustrating the synthesis of compound 43.

FIG. 14 is a diagram illustrating the synthesis of compound 46.

FIG. 15 is a diagram illustrating a prophetic synthesis of intermediate 69.

FIG. 16 is a diagram illustrating a prophetic synthesis of compound 73.

Disclosed herein are glycomimetic E-selectin antagnoists, pharmaceutical compositions comprising the same, and methods for inhibiting E-selectin mediated functions using the same. The compounds and compositions of the present disclosure may be useful for treating and/or preventing at least one disease, disorder, and/or condition that is treatable by inhibiting binding of E-selectin to one or more ligands.

The compounds of the present disclosure may have at least one improved physicochemical, pharmacological, and/or pharmacokinetic property.

In some embodiments, presented are glycomimetic E-selectin antagonists of Formula (I):

prodrugs of Formula (I), and pharmaceutically acceptable salts of any of the foregoing, wherein

R¹ is chosen from H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₁₋₈ haloalkyl, C₂₋₈ haloalkenyl, C₂₋₈ haloalkynyl,

groups, wherein n is chosen from integers ranging from 0 to 2, R⁷ is chosen from H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₄₋₁₆ cycloalkylalkyl, and —C(═O)R⁸ groups, and each R⁸ is independently chosen from H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₄₋₁₆ cycloalkylalkyl, C₆₋₁₈ aryl, and C₁₋₁₃ heteroaryl groups;

R² is chosen from C₁₋₁₂ alkyl groups substituted with at least one substituent chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups and C₂₋₁₂ alkenyl groups substituted with at least one substituent chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups, wherein each Q is independently chosen from H and pharmaceutically acceptable cations;

R³ is chosen from —OH, —OY¹, halo, —NH₂, —NHY¹, —NY¹Y², —OC(═O)Y¹, —NHC(═O)Y¹, and —NHC(═O)NHY¹ groups, wherein Y¹ and Y², which may be the same or different, are independently chosen from C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₄₋₁₆ cycloalkylalkyl, C₂₋₁₂ heterocyclyl, C₆₋₁₈ aryl, and C₁₋₁₃ heteroaryl groups, wherein Y¹ and Y² may join together along with the nitrogen atom to which they are attached to form a ring;

R⁴ is chosen from —CN, —CH₂CN, and —C(═O)Y³ groups, wherein Y³ is chosen from C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, —NHOH, —NHOCH₃, —NHCN, and —NZ¹Z² groups, wherein Z¹ and Z², which may be identical or different, are independently chosen from H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₁₋₁₂ haloalkyl, C₂₋₁₂ haloalkenyl, C₂₋₁₂ haloalkynyl, and C₇₋₁₂ arylalkyl groups, wherein Z¹ and Z² may join together along with the nitrogen atom to which they are attached to form a ring;

R⁵ is chosen from H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₁₋₁₂ haloalkyl, C₂₋₁₂ haloalkenyl, C₂₋₁₂ haloalkynyl, C₄₋₁₆ cycloalkylalkyl, and C₆₋₁₈ aryl groups;

R⁶ is chosen from —CN, C₁₋₁₂ alkyl, and C₁₋₁₂ haloalkyl groups;

L is chosen from chosen from a bond, C₆₋₁₈ aryl, and C₁₋₁₂ heteroaryl groups; and

M is chosen from linker groups;

with the proviso that when L is a bond and R² is chosen from —OH and —CO₂Q groups, then M is not a steroidal moiety.

In some embodiments, R¹ is chosen from H, C₁₋₁₂ alkyl, and C₁₋₁₂ haloalkyl groups. In some embodiments, R¹ is chosen from H and C₁₋₈ alkyl groups. In some embodiments, R¹ is H. In some embodiments, R¹ is chosen from C₁₋₆ alkyl groups. In some embodiments, R¹ is chosen from C₁₋₄ alkyl groups. In some embodiments, R¹ is chosen from methyl and ethyl. In some embodiments, R¹ is methyl. In some embodiments, R¹ is ethyl.

In some embodiments, R¹ is chosen from

In some embodiments, R¹ is chosen from

In some embodiments, R⁷ is chosen from H, C₁₋₈ alkyl, and —C(═O)R⁸ groups. In some embodiments, R⁷ is chosen from H and C₁₋₈ alkyl groups. In some embodiments, R⁷ is chosen from C₁₋₄ alkyl groups. In some embodiments, R⁷ is H.

In some embodiments, each R⁸ is independently chosen from H, C₁₋₈ alkyl, C₆₋₁₈ aryl groups, and C₁₋₁₃ heteroaryl groups. In some embodiments, at least one R⁸ is chosen from C₁₋₈ alkyl groups. In some embodiments, at least one R⁸ is chosen from C₁₋₄ alkyl groups. In some embodiments, at least one R⁸ is chosen from methyl and ethyl. In some embodiments, at least one R⁸ is H. In some embodiments, at least one R⁷ is methyl. In some embodiments, at least one R⁸ is ethyl.

In some embodiments, at least one R⁸ is chosen from

In some embodiments, R¹ is

In some embodiments, R¹ is

In some embodiments, R¹ is

In some embodiments, R² is chosen from C₁₋₁₂ alkyl groups substituted with at least one substituent chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen from C₂₋₁₂ alkenyl groups substituted with at least one substituent chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments, R² is chosen from C₁₋₈ alkyl groups substituted with at least one substituent chosen —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen from C₂₋₈ alkenyl groups substituted with at least one substituent chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments, R² is chosen from C₁₋₅ alkyl groups substituted with at least one substituent chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen from C₂₋₅ alkenyl groups substituted with at least one substituent chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments, the at least one substituent of R² is —OH. In some embodiments, the at least one substituent of R² is chosen from —OSO₃Q groups. In some embodiments, the at least one substituent of R² is chosen from —OPO₃Q₂ groups. In some embodiments, the at least one substituent of R² is chosen from —CO₂Q groups. In some embodiments, the at least one substituent of R² is chosen from —SO₃Q groups.

In some embodiments, R² is chosen from C₁₋₈ alkyl groups substituted with at least two substituents independently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen from C₂₋₈ alkenyl groups substituted with at least two substituents independently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments, R² is chosen from C₁₋₅ alkyl groups substituted with at least two substituents independently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen from C₂₋₅ alkenyl groups substituted with at least two substituents independently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments, the at least two substituents of R² are —OH. In some embodiments, the at least two substituents of R² are independently chosen from —OSO₃Q groups. In some embodiments, the at least two substituents of R² are independently chosen from —OPO₃Q₂ groups. In some embodiments, the at least two substituents of R² are independently chosen from —CO₂Q groups. In some embodiments, the at least two substituents of R² are independently chosen from —SO₃Q groups.

In some embodiments, R² is chosen from C₁₋₈ alkyl groups substituted with at least three substituents independently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen from C₂₋₈ alkenyl groups substituted with at least three substituents independently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments, R² is chosen from C₁₋₅ alkyl groups substituted with at least three substituents independently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen from C₂₋₅ alkenyl groups substituted with at least three substituents independently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments, the at least three substituents of R² are —OH. In some embodiments, the at least three substituents of R² are independently chosen from —OSO₃Q groups. In some embodiments, the at least three substituents of R² are independently chosen from —OPO₃Q₂ groups. In some embodiments, the at least three substituents of R² are independently chosen from —CO₂Q groups. In some embodiments, the at least three substituents of R² are independently chosen from —SO₃Q groups.

In some embodiments, at least one Q is H. In some embodiments, at least one Q is chosen from pharmaceutically acceptable cations. In some embodiments, at least one Q is chosen from sodium, potassium, lithium, ammonium (substituted and unsubstituted), calcium, magnesium, iron, zinc, copper, manganese, and aluminum cations. In some embodiments, at least one Q is a sodium cation. In some embodiments, at least one Q is a potassium cation. In some embodiments, at least one Q is chosen from ammonium cations.

In some embodiments, each Q is H. In some embodiments, each Q is chosen from pharmaceutically acceptable cations. In some embodiments, each Q is chosen from sodium, potassium, lithium, ammonium (substituted and unsubstituted), calcium, magnesium, iron, zinc, copper, manganese, and aluminum cations. In some embodiments, each Q is a sodium cation. In some embodiments, each Q is a potassium cation. In some embodiments, each Q is chosen from ammonium cations.

In some embodiments, R³ is chosen from —OH, —OY¹, —OC(═O)Y¹, —NH₂, and —NHC(═O)Y¹ groups, wherein Y¹ is chosen from C₁₋₈ alkyl, C₄₋₁₂ cycloalkylalkyl, C₂₋₈ heterocyclyl, C₆₋₁₂ aryl, and C₁₋₉ heteroaryl groups. In some embodiments, R³ is chosen from —OY¹ groups. In some embodiments, R³ is chosen from —OC(═O)Y¹ groups. In some embodiments, R³ is chosen from —NHC(═O)Y¹ groups. In some embodiments, R³ is —OH. In some embodiments, R³ is —NH₂.

In some embodiments, R³ is chosen from

In some embodiments, R³ is

In some embodiments, R³ is

In some embodiments, R⁴ is —CN. In some embodiments, R⁴ is —CH₂CN. In some embodiments, R⁴ is chosen from —C(═O)Y³ groups, wherein Y³ is chosen from —OZ¹, —NHOH, —NHOCH₃, and —NZ¹Z² groups. In some embodiments, R⁴ is chosen from —C(═O)OZ¹ groups. In some embodiments, R⁴ is chosen from —C(═O)NZ¹Z² groups. In some embodiments, Z¹ and Z², which may be identical or different, are independently chosen from H, C₁₋₈ alkyl, C₁₋₈ haloalkyl, and C₇₋₁₂ arylalkyl groups. In some embodiments, at least one of Z¹ and Z² is H. In some embodiments, each of Z¹ and Z² is H. In some embodiments, at least one of Z¹ and Z² is methyl. In some embodiments, each of Z¹ and Z² is methyl. In some embodiments, at least one of Z¹ and Z² is ethyl. In some embodiments, each of Z¹ and Z² is ethyl. In some embodiments, Z¹ is H and Z² is methyl. In some embodiments, Z¹ and Z² join together along with the nitrogen atom to which they are attached to form a ring.

In some embodiments, R⁴ is chosen from

In some embodiments, R⁴ is chosen from

In some embodiments, R⁴ is

In some embodiments, R⁴ is

In some embodiments, R⁴ is

In some embodiments, R⁴ is

In some embodiments, R⁵ is chosen from C₁₋₁₂ alkyl groups. In some embodiments, R⁵ is chosen from C₁₋₈ alkyl groups. In some embodiments, R⁵ is chosen from C₁₋₁₂ haloalkyl groups. In some embodiments, R⁵ is chosen from C₁₋₈ haloalkyl groups. In some embodiments, R⁵ is chosen from C₄₋₁₆ cycloalkylalkyl groups. In some embodiments, R⁵ is chosen from C₄₋₈ cycloalkylalkyl groups. In some embodiments, R⁵ is chosen from propyl, cyclopropylmethyl, and cyclohexylmethyl. In some embodiments, R⁵ is propyl. In some embodiments, R⁵ is cyclopropylmethyl. In some embodiments, R⁵ is cyclohexylmethyl.

In some embodiments, R⁶ is chosen from C₁₋₁₂ alkyl groups. In some embodiments, R⁶ is chosen from C₁₋₄ alkyl groups. In some embodiments, R⁶ is chosen from C₁₋₁₂ haloalkyl groups. In some embodiments, R⁶ is chosen from C₁₋₄ haloalkyl groups. In some embodiments, R⁶ is chosen from halomethyl groups. In some embodiments, R⁶ is CF₃. In some embodiments, R⁶ is CH₃. In some embodiments, R⁶ is CN.

In some embodiments, L is a bond. In some embodiments L is chosen from C₆₋₁₈ aryl groups. In some embodiments L is chosen from C₁₋₁₂ heteroaryl groups.

In some embodiments, L is

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, linker groups (M) may be chosen from groups comprising spacer groups, such spacer groups as, for example, —(CH₂)_(t)— and —O(CH₂)_(t)—, wherein t is chosen from integers ranging from 1 to 20. Other non-limiting examples of spacer groups include carbonyl groups and carbonyl-containing groups such as, for example, amide groups. A non-limiting example of a spacer group is:

In some embodiments, the linker group (M) is chosen from:

In some embodiments, the linker group (M) is chosen from polyethylene glycols (PEGS), —C(═O)NH(CH₂)_(v)O—, —C(═O)NH(CH₂)_(v)NHC(═O)—, —C(═O)NHC(═O)(CH₂)NH— and —C(═O)NH(CH₂)_(v)C(═O)NH— groups, wherein v is chosen from integers ranging from 2 to 20. In some embodiments, v is chosen from integers ranging from 2 to 4. In some embodiments, v is 2. In some embodiments, v is 3. In some embodiments, v is 4.

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, the linker group (M) is:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

In some embodiments, at least one compound is chosen from compounds having the following Formulae:

Also provided are pharmaceutical compositions comprising at least one compound of Formula (I). Such pharmaceutical compositions are described in greater detail herein. These compounds and compositions may be used in the methods described herein.

In some embodiments, a method for treating and/or preventing at least one disease, disorder, and/or condition where inhibition of E-selectin mediated functions may be useful is disclosed, the method comprising administering at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a method for treating and/or preventing at least one inflammatory disease, disorder, and/or condition in which the adhesion and/or migration of cells occurs in the disease, disorder, and/or condition is disclosed, the method comprising administering at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a method for inhibiting adhesion of a cancer cell that expresses a ligand of E-selectin to an endothelial cell expressing E-selectin on the cell surface of the endothelial cell is disclosed, the method comprising contacting the endothelial cell and at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) such that the at least one compound of Formula (I) interacts with E-selectin on the endothelial cell, thereby inhibiting binding of the cancer cell to the endothelial cell. In some embodiments, the endothelial cell is present in the bone marrow.

In some embodiment, a method for treating and/or preventing a cancer is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I). In some embodiments, at least one compound of Formula (I) and/or pharmaceutical composition comprising at least one compound of Formula (I) may be administered in conjunction with (i.e., as an adjunct therapy, which is also called adjunctive therapy) chemotherapy and/or radiotherapy.

The chemotherapy and/or radiotherapy may be referred to as the primary anti-tumor or anti-cancer therapy that is being administered to the subject to treat the particular cancer. In some embodiments, a method for reducing (i.e., inhibiting, diminishing) chemosensitivity and/or radiosensitivity of hematopoietic stem cells (HSC) to the chemotherapeutic drug(s) and/or radiotherapy, respectively, is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a method for enhancing (i.e., promoting) survival of hematopoietic stem cells is provided, the method comprising administering to a subject in need thereof at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a method for decreasing the likelihood of occurrence of metastasis of cancer cells (also called tumor cells herein) in a subject who is in need thereof is disclosed, the method comprising administering an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a method for treatment and/or prevention of at least one cancer in which the cancer cells may leave the primary site is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I). A primary site may be, for example, solid tissue (e.g., breast or prostate) or the bloodstream.

In some embodiments, a method for treatment and/or prevention of at least one cancer in which it is desirable to mobilize cancer cells from a site into the bloodstream and/or retain the cancer cells in the bloodstream is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a method for decreasing the likelihood of occurrence of infiltration of cancer cells into bone marrow is disclosed, the method comprises administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a method for releasing cells into circulating blood and enhancing retention of the cells in the blood is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I). In some embodiments, the method further includes collecting the released cells. In some embodiments, collecting the released cells utilizes apheresis. In some embodiments, the released cells are stem cells (e.g., bone marrow progenitor cells). In some embodiments, G-CSF is administered to the individual.

In some embodiments, a method for treating and/or preventing thrombosis is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a method for treating and/or preventing mucositis is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a method for treating and/or preventing one cardiovascular disease, disorder and/or condition is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a method for treatment and/or prevention of atherosclerosis is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

In some embodiments, a compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) may be used for the preparation and/or manufacture of a medicament for use in treating and/or preventing at least one of the diseases, disorders, and/or conditions described herein.

Whenever a term in the specification is identified as a range (e.g., C₁₋₄ alkyl) or “ranging from”, the range independently discloses and includes each element of the range. As a non-limiting example, C₁₋₄ alkyl groups includes, independently, C₁ alkyl groups, C₂ alkyl groups, C₃ alkyl groups, and C₄ alkyl groups. As another non-limiting example, “n is an integer ranging from 0 to 2” includes, independently, 0, 1, and 2.

The term “at least one” refers to one or more, such as one, two, etc. For example, the term “at least one C₁₋₄ alkyl group” refers to one or more C₁₋₄ alkyl groups, such as one C₁₋₄ alkyl group, two C₁₋₄ alkyl groups, etc.

The term “alkyl” includes saturated straight, branched, and cyclic (also identified as cycloalkyl), primary, secondary, and tertiary hydrocarbon groups. Non-limiting examples of alkyl groups include methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, secbutyl, isobutyl, tertbutyl, cyclobutyl, 1-methylbutyl, 1,1-dimethylpropyl, pentyl, cyclopentyl, isopentyl, neopentyl, cyclopentyl, hexyl, isohexyl, and cyclohexyl. Unless stated otherwise specifically in the specification, an alkyl group may be optionally substituted.

The term “alkenyl” includes straight, branched, and cyclic hydrocarbon groups comprising at least one double bond. The double bond of an alkenyl group can be unconjugated or conjugated with another unsaturated group. Non-limiting examples of alkenyl groups include vinyl, allyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, 2-ethylhexenyl, and cyclopent-1-en-1-yl. Unless stated otherwise specifically in the specification, an alkenyl group may be optionally substituted.

The term “alkynyl” includes straight and branched hydrocarbon groups comprising at least one triple bond. The triple bond of an alkynyl group can be unconjugated or conjugated with another unsaturated group. Non-limiting examples of alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, and hexynyl. Unless stated otherwise specifically in the specification, an alkynyl group may be optionally substituted.

The term “aryl” includes hydrocarbon ring system groups comprising at least 6 carbon atoms and at least one aromatic ring. The aryl group may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems. Non-limiting examples of aryl groups include aryl groups derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Unless stated otherwise specifically in the specification, an aryl group may be optionally substituted.

The terms “E-selectin antagonist” and “E-selectin inhibitor” are used interchangeably herein, and include inhibitors of E-selectin only, as well as inhibitors of E-selectin and either P-selectin or L-selectin, and inhibitors of E-selectin, P-selectin, and L-selectin.

The term “glycomimetic” includes any naturally occurring or non-naturally occurring carbohydrate compound in which at least one substituent has been replaced, or at least one ring has been modified (e.g., substitution of carbon for a ring oxygen), to yield a compound that is not fully carbohydrate.

The term “halo” or “halogen” includes fluoro, chloro, bromo, and iodo.

The term “haloalkyl” includes alkyl groups, as defined herein, substituted by at least one halogen, as defined herein. Non-limiting examples of haloalkyl groups include trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, and 1,2-dibromoethyl. A “fluoroalkyl” is a haloalkyl wherein at least one halogen is fluoro. Unless stated otherwise specifically in the specification, a haloalkyl group may be optionally substituted.

The term “haloalkenyl” includes alkenyl groups, as defined herein, substituted by at least one halogen, as defined herein. Non-limiting examples of haloalkenyl groups include fluoroethenyl, 1,2-difluoroethenyl, 3-bromo-2-fluoropropenyl, and 1,2-dibromoethenyl. A “fluoroalkenyl” is a haloalkenyl substituted with at least one fluoro group. Unless stated otherwise specifically in the specification, a haloalkenyl group may be optionally substituted.

The term “haloalkynyl” includes alkynyl groups, as defined herein, substituted by at least one halogen, as defined herein. Non-limiting examples include fluoroethynyl, 1,2-difluoroethynyl, 3-bromo-2-fluoropropynyl, and 1,2-dibromoethynyl. A “fluoroalkynyl” is a haloalkynyl wherein at least one halogen is fluoro. Unless stated otherwise specifically in the specification, a haloalkynyl group may be optionally substituted.

The term “heterocyclyl” or “heterocyclic ring” includes 3- to 24-membered saturated or partially unsaturated non-aromatic ring groups comprising 2 to 23 ring carbon atoms and 1 to 8 ring heteroatom(s) each independently chosen from N, O, and S. Unless stated otherwise specifically in the specification, the heterocyclyl groups may be monocyclic, bicyclic, tricyclic or tetracyclic ring systems, which may include fused or bridged ring systems, and may be partially or fully saturated; any nitrogen, carbon or sulfur atom(s) in the heterocyclyl group may be optionally oxidized; any nitrogen atom in the heterocyclyl group may be optionally quaternized; and the heterocyclyl group Non-limiting examples of heterocyclic ring include dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in the specification, a heterocyclyl group may be optionally substituted.

The term “heteroaryl” includes 5- to 14-membered ring groups comprising 1 to 13 ring carbon atoms and 1 to 6 ring heteroatom(s) each independently chosen from N, O, and S, and at least one aromatic ring. Unless stated otherwise specifically in the specification, the heteroaryl group may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl radical may be optionally oxidized; the nitrogen atom may be optionally quaternized. Non-limiting examples include azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e. thienyl). Unless stated otherwise specifically in the specification, a heteroaryl group may be optionally substituted.

The term “pharmaceutically acceptable salts” includes both acid and base addition salts. Non-limiting examples of pharmaceutically acceptable acid addition salts include chlorides, bromides, sulfates, nitrates, phosphates, sulfonates, methane sulfonates, formates, tartrates, maleates, citrates, benzoates, salicylates, and ascorbates. Non-limiting examples of pharmaceutically acceptable base addition salts include sodium, potassium, lithium, ammonium (substituted and unsubstituted), calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Pharmaceutically acceptable salts may, for example, be obtained using standard procedures well known in the field of pharmaceuticals.

The term “prodrug” includes compounds that may be converted, for example, under physiological conditions or by solvolysis, to a biologically active compound described herein. Thus, the term “prodrug” includes metabolic precursors of compounds described herein that are pharmaceutically acceptable. A discussion of prodrugs can be found, for example, in Higuchi, T., et al., “Pro-drugs as Novel Delivery Systems,” A.C.S. Symposium Series, Vol. 14, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987. The term “prodrug” also includes covalently bonded carriers that release the active compound(s) as described herein in vivo when such prodrug is administered to a subject. Non-limiting examples of prodrugs include ester and amide derivatives of hydroxy, carboxy, mercapto and amino functional groups in the compounds described herein.

The term “substituted” includes the situation where, in any of the above groups, at least one hydrogen atom is replaced by a non-hydrogen atom such as, for example, a halogen atom such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxyl groups, alkoxy groups, and ester groups; a sulfur atom in groups such as thiol groups, thioalkyl groups, sulfone groups, sulfonyl groups, and sulfoxide groups; a nitrogen atom in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; a silicon atom in groups such as trialkylsilyl groups, dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl groups; and other heteroatoms in various other groups. “Substituted” also includes the situation where, in any of the above groups, at least one hydrogen atom is replaced by a higher-order bond (e.g., a double- or triple-bond) to a heteroatom such as oxygen in oxo, carbonyl, carboxyl, and ester groups; and nitrogen in groups such as imines, oximes, hydrazones, and nitriles.

The present disclosure includes within its scope all the possible geometric isomers, e.g., Z and E isomers (cis and trans isomers), of the compounds as well as all the possible optical isomers, e.g., diastereomers and enantiomers, of the compounds. Furthermore, the present disclosure includes in its scope both the individual isomers and any mixtures thereof, e.g., racemic mixtures. The individual isomers may be obtained using the corresponding isomeric forms of the starting material or they may be separated after the preparation of the end compound according to conventional separation methods. For the separation of optical isomers, e.g., enantiomers, from the mixture thereof conventional resolution methods, e.g., fractional crystallization, may be used.

The present disclosure includes within its scope all possible tautomers. Furthermore, the present disclosure includes in its scope both the individual tautomers and any mixtures thereof.

Compounds of Formula (I) may be prepared as shown in, for example, FIGS. 5, 7, 9-11, and 13-14. It is understood that one of ordinary skill in the art may be able to make these compounds by similar methods or by combining other methods known to one of ordinary skill in the art. It is also understood that one of ordinary skill in the art would be able to make other compounds of Formula (I) not specifically illustrated herein by using appropriate starting components and modifying the parameters of the synthesis as needed (e.g., see FIG. 16). In general, starting components may be obtained from sources such as Sigma Aldrich, Alfa Aesar, Maybridge, Matrix Scientific, TCI, and Fluorochem USA, etc. and/or synthesized according to sources known to those of ordinary skill in the art (see, for example, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th edition (Wiley, December 2000)) and/or prepared as described herein.

It will also be appreciated by those skilled in the art that in the processes described herein the functional groups of intermediate compounds may need to be protected by suitable protecting groups, even if not specifically described. Such functional groups include hydroxy, amino, mercapto, and carboxylic acid. Suitable protecting groups for hydroxy include but are not limited to trialkylsilyl or diarylalkylsilyl (for example, t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino, amidino and guanidino include but are not limited to t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for mercapto include but are not limited to —C(O)R″ (where R″ is alkyl, aryl or arylalkyl), p-methoxybenzyl, trityl and the like. Suitable protecting groups for carboxylic acid include but are not limited to alkyl, aryl or arylalkyl esters. Protecting groups may be added or removed in accordance with standard techniques, which are known to one skilled in the art and as described herein. The use of protecting groups is described in detail in Green, T. W. and P. G. M. Wutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley. As one of skill in the art would appreciate, the protecting group may also be a polymer resin such as a Wang resin, Rink resin or a 2-chlorotrityl-chloride resin.

Analogous reactants to those described herein may be identified through the indices of known chemicals prepared by the Chemical Abstract Service of the American Chemical Society, which are available in most public and university libraries, as well as through on-line databases (the American Chemical Society, Washington, D.C., may be contacted for more details). Chemicals that are known but not commercially available in catalogs may be prepared by custom chemical synthesis houses, where many of the standard chemical supply houses (e.g., those listed above) provide custom synthesis services. A reference for the preparation and selection of pharmaceutical salts of the present disclosure is P. H. Stahl & C. G. Wermuth “Handbook of Pharmaceutical Salts,” Verlag Helvetica Chimica Acta, Zurich, 2002.

Methods known to one of ordinary skill in the art may be identified through various reference books, articles, and databases. Suitable reference books and treatise that detail the synthesis of reactants useful in the preparation of compounds of the present disclosure, or provide references to articles that describe the preparation, include for example, “Synthetic Organic Chemistry,” John Wiley & Sons, Inc., New York; S. R. Sandler et al., “Organic Functional Group Preparations,” 2nd Ed., Academic Press, New York, 1983; H. O. House, “Modern Synthetic Reactions”, 2nd Ed., W. A. Benjamin, Inc. Menlo Park, Calif. 1972; T. L. Gilchrist, “Heterocyclic Chemistry”, 2nd Ed., John Wiley & Sons, New York, 1992; J. March, “Advanced Organic Chemistry: Reactions, Mechanisms and Structure,” 4th Ed., Wiley-Interscience, New York, 1992. Additional suitable reference books and treatise that detail the synthesis of reactants useful in the preparation of compounds of the present disclosure, or provide references to articles that describe the preparation, include for example, Fuhrhop, J. and Penzlin G. “Organic Synthesis: Concepts, Methods, Starting Materials”, Second, Revised and Enlarged Edition (1994) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R. V. “Organic Chemistry, An Intermediate Text” (1996) Oxford University Press, ISBN 0-19-509618-5; Larock, R. C. “Comprehensive Organic Transformations: A Guide to Functional Group Preparations” 2nd Edition (1999) Wiley-VCH, ISBN: 0-471-19031-4; March, J. “Advanced Organic Chemistry: Reactions, Mechanisms, and Structure” 4th Edition (1992) John Wiley & Sons, ISBN: 0-471-60180-2; Otera, J. (editor) “Modern Carbonyl Chemistry” (2000) Wiley-VCH, ISBN: 3-527-29871-1; Patai, S. “Patai's 1992 Guide to the Chemistry of Functional Groups” (1992) Interscience ISBN: 0-471-93022-9; Quin, L. D. et al. “A Guide to Organophosphorus Chemistry” (2000) Wiley-Interscience, ISBN: 0-471-31824-8; Solomons, T. W. G. “Organic Chemistry” 7th Edition (2000) John Wiley & Sons, ISBN: 0-471-19095-0; Stowell, J. C., “Intermediate Organic Chemistry” 2nd Edition (1993) Wiley-Interscience, ISBN: 0-471-57456-2; “Industrial Organic Chemicals: Starting Materials and Intermediates: An Ullmann's Encyclopedia” (1999) John Wiley & Sons, ISBN: 3-527-29645-X, in 8 volumes; “Organic Reactions” (1942-2000) John Wiley & Sons, in over 55 volumes; and “Chemistry of Functional Groups” John Wiley & Sons, in 73 volumes.

Biological activity of a compound described herein may be determined, for example, by performing at least one in vitro and/or in vivo study routinely practiced in the art and described herein or in the art. In vitro assays include without limitation binding assays, immunoassays, competitive binding assays, and cell based activity assays.

An inhibition assay may be used to screen for antagonists of E-selectin. For example, an assay may be performed to characterize the capability of a compound described herein to inhibit (i.e., reduce, block, decrease, or prevent in a statistically or biologically significant manner) interaction of E-selectin with sLe^(a) or sLe^(x). The inhibition assay may be a competitive binding assay, which allows the determination of IC₅₀ values. By way of example, E-selectin/Ig chimera may be immobilized onto a matrix (e.g., a multi-well plate, which may be made from a polymer, such as polystyrene; a test tube, and the like); a composition may be added to reduce nonspecific binding (e.g., a composition comprising non-fat dried milk or bovine serum albumin or other blocking buffer routinely used by a person skilled in the art); the immobilized E-selectin may be contacted with the candidate compound in the presence of sLe^(a) comprising a reporter group under conditions and for a time sufficient to permit sLe^(a) to bind to the immobilized E-selectin; the immobilized E-selectin may be washed; and the amount of sLe^(a) bound to immobilized E-selectin may be detected. Variations of such steps can be readily and routinely accomplished by a person of ordinary skill in the art.

Conditions for a particular assay include temperature, buffers (including salts, cations, media), and other components that maintain the integrity of any cell used in the assay and the compound, which a person of ordinary skill in the art will be familiar and/or which can be readily determined. A person of ordinary skill in the art also readily appreciates that appropriate controls can be designed and included when performing the in vitro methods and in vivo methods described herein.

The source of a compound that is characterized by at least one assay and techniques described herein and in the art may be a biological sample that is obtained from a subject who has been treated with the compound. The cells that may be used in the assay may also be provided in a biological sample. A “biological sample” may include a sample from a subject, and may be a blood sample (from which serum or plasma may be prepared), a biopsy specimen, one or more body fluids (e.g., lung lavage, ascites, mucosal washings, synovial fluid, urine), bone marrow, lymph nodes, tissue explant, organ culture, or any other tissue or cell preparation from the subject or a biological source. A biological sample may further include a tissue or cell preparation in which the morphological integrity or physical state has been disrupted, for example, by dissection, dissociation, solubilization, fractionation, homogenization, biochemical or chemical extraction, pulverization, lyophilization, sonication, or any other means for processing a sample derived from a subject or biological source. In some embodiments, the subject or biological source may be a human or non-human animal, a primary cell culture (e.g., immune cells), or culture adapted cell line, including but not limited to, genetically engineered cell lines that may contain chromosomally integrated or episomal recombinant nucleic acid sequences, immortalized or immortalizable cell lines, somatic cell hybrid cell lines, differentiated or differentiatable cell lines, transformed cell lines, and the like.

As described herein, methods for characterizing E-selectin include animal model studies. Non-limiting examples of animal models for liquid cancers used in the art include multiple myeloma (see, e.g., DeWeerdt, Nature 480:S38-S39 (15 Dec. 2011) doi:10.1038/480S38a; Published online 14 Dec. 2011; Mitsiades et al., Clin. Cancer Res. 2009 15:1210021 (2009)); acute myeloid leukemia (AML) (Zuber et al., Genes Dev. 2009 Apr. 1; 23(7): 877-889). Animal models for acute lymphoblastic leukemia (ALL) have been used by persons of ordinary skill in the art for more than two decades. Numerous exemplary animal models for solid tumor cancers are routinely used and are well known to persons of ordinary skill in the art.

The compounds of the present disclosure and the pharmaceutical compositions comprising at least one of such compounds may be useful in methods for treating and/or preventing a disease or disorder that is treatable by inhibiting at least one activity of E-selectin (and/or inhibiting binding of E-selectin to ligand(s), which in turn inhibits a biological activity).

The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may be useful in methods for treating and/or preventing at least one inflammatory disease. Inflammation comprises reaction of vascularized living tissue to injury. By way of example, although E-selectin mediated cell adhesion may be important to the body's anti-infective immune response, in other circumstances, E-selectin mediated cell adhesion may be undesirable or excessive, resulting in tissue damage and/or scarring instead of repair. For example, many pathologies (such as autoimmune and inflammatory diseases, shock and reperfusion injuries) involve abnormal adhesion of white blood cells. Therefore, inflammation affects blood vessels and adjacent tissues in response to an injury or abnormal stimulation by a physical, chemical, or biological agent. Examples of inflammatory diseases, disorders, or conditions include, without limitation, dermatitis, chronic eczema, psoriasis, multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, graft versus host disease, sepsis, diabetes, atherosclerosis, Sjogren's syndrome, progressive systemic sclerosis, scleroderma, acute coronary syndrome, ischemic reperfusion, Crohn's disease, inflammatory bowel disease, endometriosis, glomerulonephritis, myasthenia gravis, idiopathic pulmonary fibrosis, asthma, allergic reaction, acute respiratory distress syndrome (ARDS) or other acute leukocyte-mediated lung injury, vasculitis, or inflammatory autoimmune myositis. Other diseases and disorders for which the compounds described herein may be useful for treating and/or preventing include hyperactive coronary circulation, microbial infection, cancer metastasis, thrombosis, wounds, burns, spinal cord damage, digestive tract mucous membrane disorders (e.g., gastritis, ulcers), osteoporosis, osteoarthritis, septic shock, traumatic shock, stroke, nephritis, atopic dermatitis, frostbite injury, adult dyspnoea syndrome, ulcerative colitis, diabetes and reperfusion injury following ischemic episodes, prevention of restenosis associated with vascular stenting, and for undesirable angiogenesis, for example, angiogenesis associated with tumor growth.

As discussed in detail herein, a disease or disorder to be treated or prevented is a cancer and related metastasis and includes cancers that comprise solid tumor(s) and cancers that comprise liquid tumor(s). The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may be useful in methods for preventing and/or treating cancer. In some embodiments, the at least one compound may be used for treating and/or preventing metastasis and/or for inhibiting (slowing, retarding, or preventing) metastasis of cancer cells.

In some embodiments, at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) is administered to a cancer patient in remission. In some embodiments, the at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) is administered as a cancer vaccine to stimulate marrow infiltrating lymphocytes (“MILs”) in a cancer patient or cancer survivor to prevent relapse.

In some embodiments, a method of treating cancer and/or preventing a cancer relapse is disclosed, wherein the method comprises administering to a patient in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I), wherein the amount of compound of Formula (I) administered is sufficient to mobilize MILs of the patient into the peripheral blood.

In some embodiments, a method of treating cancer and/or preventing a cancer relapse is provided comprising administering to a donor patient at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) in an amount of sufficient to mobilize MILs of the patient out of the marrow (e.g., into the peripheral blood), recovering MILS (e.g., recovering them from the peripheral blood), and transplanting at least a portion of the MIL cell population to the donor patient or another patient. In some embodiments, the MIL cell population is expanded ex vivo before transplantation.

In some embodiments, a method of preventing cancer is provided comprising administering to a donor patient at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) in an amount sufficient to mobilize MILs of the patient out of the bone marrow (e.g., into the peripheral blood), recovering MILs (e.g., recovering them from the peripheral blood), and transplanting at least a portion of MIL cell population to a subject (e.g., a non-cancer patient, a patient suffering from a different form or type of cancer than the donor patient, etc.). In some embodiments, the MIL cell population is expanded ex vivo before transplantation.

In some embodiments, the compounds of present disclosure and pharmaceutical compositions comprising at least one such compound may be used for decreasing (i.e., reducing) the likelihood of occurrence of metastasis of cancer cells in an individual (i.e., subject, patient) who is in need thereof. The compounds of the present disclosure and compositions comprising at least one such compound may be used for decreasing (i.e., reducing) the likelihood of occurrence of infiltration of cancer cells into bone marrow in an individual who is in need thereof. The individuals (or subjects) in need of such treatments include subjects who have been diagnosed with a cancer, which includes cancers that comprise solid tumor(s) and cancers that comprise liquid tumor(s).

Non-limiting examples of cancers include colorectal cancers, liver cancers, gastric cancers, lung cancers, brain cancers, kidney cancers, bladder cancers, thyroid cancers, prostate cancers, ovarian cancers, cervical cancers, uterine cancers, endometrial cancers, melanomas, breast cancers, and pancreatic cancers. Liquid tumors can occur in the blood, bone marrow, the soft, sponge-like tissue in the center of most bones, and lymph nodes and include leukemias (e.g., AML, ALL, CLL, and CML), lymphomas, and myelomas (e.g., multiple myeloma). Lymphomas include Hodgkin lymphoma, which is marked by the presence of a type of cell called the Reed-Sternberg cell, and non-Hodgkin lymphomas, which includes a large, diverse group of cancers of immune system cells. Non-Hodgkin lymphomas can be further divided into cancers that have an indolent (slow-growing) course and those that have an aggressive (fast-growing) course, and which subtypes respond to treatment differently.

The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may be administered as an adjunct therapy to chemotherapy and/or radiotherapy, which is/are being delivered to the subject as primary therapy for treating the cancer. The chemotherapy and/or radiotherapy that may be administered depend upon several factors including the type of cancer, location of the tumor(s), stage of the cancer, age and gender and general health status of the subject. A person of ordinary skill in the medical art can readily determine the appropriate chemotherapy regimen and/or radiotherapy regimen for the subject in need. The person of ordinary skill in the medical art can also determine, with the aid of preclinical and clinical studies, when the compound of the present disclosure or pharmaceutical composition comprising at least one such compound should be administered to the subject, that is whether the compound or composition is administered prior to, concurrent with, or subsequent to a cycle of the primary chemotherapy or radiation treatment.

Also provided herein is a method for inhibiting adhesion of a tumor cell that expresses a ligand of E-selectin to an endothelial cell expressing E-selectin on its cell surface, which method comprises contacting the endothelial cell with at least one compound of the present disclosure or pharmaceutical compositions comprising at least one such compound, thereby permitting the compound to interact with E-selectin on the endothelial cell surface and inhibiting binding of the tumor cell to the endothelial cell. Without wishing to be bound by theory, inhibiting adhesion of tumor cells to endothelial cells may reduce in a significant manner, the capability of the tumor cells to extravasate into other organs, blood vessels, lymph, or bone marrow and thereby reduce, decrease, or inhibit, or slow the progression of the cancer, including reducing, decreasing, inhibiting, or slowing metastasis.

In some embodiments, a method for inhibiting adhesion of metastasized tumor cells is disclosed, the method comprising administering at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I).

As described herein, at least one of the compounds of the present disclosure or pharmaceutical compositions comprising at least one such compound may be administered in combination with at least one additional anti-cancer agent. Chemotherapy may comprise one or more chemotherapeutic agents. For example, chemotherapy agents, radiotherapeutic agents, inhibitors of phosphoinositide-3 kinase (PI3K), and inhibitors of VEGF may be used in combination with a compound of Formula (I) described herein. Non-limiting examples of inhibitors of PI3K include the compound named by Exelixis as “XL499.” Non-limiting examples of VEGF inhibitors include the compound called “cabo” (previously known as XL184). Many other chemotherapeutics are small organic molecules. As understood by a person of ordinary skill in the art, chemotherapy may also refer to a combination of two or more chemotherapeutic molecules that are administered coordinately and which may be referred to as combination chemotherapy. Numerous chemotherapeutic drugs are used in the oncology art and include, for example, alkylating agents; antimetabolites; anthracyclines, plant alkaloids; and topoisomerase inhibitors.

The compounds of the present disclosure or pharmaceutical compositions comprising at least one such compound may function independently from the anti-cancer agent or may function in coordination with the anti-cancer agent, e.g., by enhancing effectiveness of the anti-cancer agent or vice versa. Accordingly, provided herein are methods for enhancing (i.e., enhancing, promoting, improving the likelihood of, enhancing in a statistically or biologically significant manner) and/or maintaining survival of hematopoietic stem cells (HSC) in a subject who is treated with and/or will be treated with a chemotherapeutic drug(s) and/or radioactive therapy, respectively, comprising administering at least one compound of Formula (I) as described herein. In some embodiments, the subject receives and/or will receive both chemotherapy and radiation therapy. Also, provided herein is a method for reducing (i.e., reducing, inhibiting, diminishing in a statistically or biologically significant manner) chemosensitivity and/or radiosensitivity of hematopoietic stem cells (HSC) to the chemotherapeutic drug(s) and/or radioactive therapy, respectively, in a subject. Because repeated cycles of chemotherapy and radiotherapy often diminish the ability of HSCs to recover and replenish bone marrow, the glycomimetic compounds described herein may be useful for subjects who will receive more than one cycle, such as at least two, three, four or more cycles, of chemotherapy and/or radiotherapy. HSCs reside in the bone marrow and generate the cells that are needed to replenish the immune system and the blood. Anatomically, bone marrow comprises a vascular niche that is adjacent to bone endothelial sinuses (see, e.g., Kiel et al., Cell 121:1109-21 (2005); Sugiyama et al., Immunity 25:977-88 (2006); Mendez-Ferrer et al., Nature 466:829-34 (2010); Butler et al., Cell Stem Cell 6:251-64 (2010)). A recent study describes that E-selectin promotes HSC proliferation and is an important component of the vascular niche (see, e.g., Winkler et al., Nature Medicine published online 21 Oct. 2012; doi:10.1038/nm.2969). Deletion or inhibition of E-selectin enhanced HSC survival in mice that were treated with chemotherapeutic agents or radiotherapy and accelerated blood neutrophil recovery (see, e.g., Winkler et al., supra).

In addition, the administration of at least one compound of the present disclosure or pharmaceutical composition comprising at least one such compounds may be in conjunction with one or more other therapies, e.g., for reducing toxicities of therapy. For example, at least one palliative agent to counteract (at least in part) a side effect of a therapy (e.g., anti-cancer therapy) may be administered. Agents (chemical or biological) that promote recovery, or counteract side effects of administration of antibiotics or corticosteroids, are examples of such palliative agents. At least one compound described herein may be administered before, after, or concurrently with administration of at least one additional anti-cancer agent or at least one palliative agent to reduce a side effect of therapy. When administration is concurrent, the combination may be administered from a single container or two (or more) separate containers.

Cancer cells (also called herein tumor cells) that may be prevented (i.e., inhibited, slowed) from metastasizing, from adhering to an endothelial cell, or from infiltrating bone marrow include cells of solid tumors and liquid tumors (including hematological malignancies). Examples of solid tumors are described herein and include colorectal cancer, liver cancer, gastric cancer, lung cancer, brain cancer, kidney cancer, bladder cancer, thyroid cancer, prostate cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, melanoma, breast cancer, and pancreatic cancer. Liquid tumors occur in the blood, bone marrow, and lymph nodes and include leukemia (e.g., AML, ALL, CLL, and CML), lymphoma (e.g., Hodgkin lymphoma and non-Hodgkin lymphoma), and myeloma (e.g., multiple myeloma). As used herein, the term cancer cells include mature, progenitor, and cancer stem cells.

Bones are a common location for cancer to infiltrate once leaving the primary tumor location. Once cancer resides in bone, it is frequently a cause of pain to the individual. In addition, if the particular bone affected is a source for production of blood cells in the bone marrow, the individual may develop a variety of blood cell related disorders. Breast and prostate cancer are examples of solid tumors that migrate to bones. Acute myelogenous leukemia (AML) and multiple myeloma (MM) are examples of liquid tumors that migrate to bones. Cancer cells that migrate to bone will typically migrate to the endosteal region of the bone marrow. Once cancer cells have infiltrated into the marrow, the cells become quiescent and are protected from chemotherapy. The compounds of the present disclosure may block infiltration of disseminated cancer cells into bone marrow. A variety of subjects may benefit from treatment with the compounds. Examples of such subjects include individuals with a cancer type having a propensity to migrate to bone where the tumor is still localized or the tumor is disseminated but not yet infiltrated bone, or where individuals with such a cancer type are in remission.

The cancer patient population most likely to respond to treatment using antagonists of E-selectin (e.g., compounds of Formula (I)) described herein can be identified based on the mechanisms of action of E-selectin. For example, patients may be selected that express a highly active E-selectin as determined by the genetic polymorphism for E-selectin of S128R (Alessandro et al., Int. J. Cancer 121:528-535, 2007). In addition, patients for treatment by the compounds described herein may also selected based on elevated expression of the E-selectin binding ligands (sialyl Le^(a) and sialyl Le^(x)) as determined by antibodies directed against cancer-associated antigens CA-19-9 (Zheng et al., World J. Gastroenterol. 7:431-434, 2001) and CD65. In addition, antibodies HECA-452 and FH-6 which recognize similar carbohydrate ligands of E-selectin may also be used in a diagnostic assay to select the cancer patient population most likely to respond to this treatment.

The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may be useful in methods for mobilizing cells from the bone marrow to the peripheral vasculature and tissues. As discussed herein, in some embodiments, the compounds and compositions are useful for mobilizing hematopoietic cells, including hematopoietic stem cells and hematopoietic progenitor cells. In some embodiments, the compounds act as mobilizing agents of normal blood cell types. In some embodiments, the agents are used in methods for mobilizing mature white blood cells (which may also be called leukocytes herein), such as granulocytes (e.g., neutrophils, eosinophils, basophils), lymphocytes, and monocytes from the bone marrow or other immune cell compartments such as the spleen and liver. Methods are also provided for using the compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound in methods for mobilizing tumor cells from the bone marrow. The tumor cells may be malignant cells (e.g., tumor cells that are metastatic cancer cells, or highly invasive tumor cells) in cancers. These tumor cells may be of hematopoietic origin or may be malignant cells of another origin residing in the bone.

In some embodiments, the methods using the compounds described herein are useful for mobilizing hematopoietic cells, such as hematopoietic stem cells and progenitor cells and leukocytes (including granulocytes such as neutrophils), which are collected (i.e., harvested, obtained) from the subject receiving a compound of Formula (I) and at a later time are administered back into the same subject (autologous donor) or administered to a different subject (allogeneic donor). Hematopoietic stem cell replacement and hematopoietic stem cell transplantation have been successfully used for treating a number of diseases (including cancers) as described herein and in the art. By way of example, stem cell replacement therapy or transplantation follows myeloablation of a subject, such as occurs with administration of high dose chemotherapy and/or radiotherapy. Desirably, an allogeneic donor shares sufficient HLA antigens with the recipient/subject to minimize the risk of host versus graft disease in the recipient (i.e., the subject receiving the hematopoietic stem cell transplant). Obtaining the hematopoietic cells from the donor subject (autologous or allogeneic) is performed by apheresis or leukapheresis. HLA typing of a potential donor and the recipient and apheresis or leukapheresis are methods routinely practiced in the clinical art.

By way of non-limiting example, autologous or allogenic hematopoietic stem cells and progenitors cells may be used for treating a recipient subject who has certain cancers, such as Hodgkin lymphoma, non-Hodgkin lymphoma, or multiple myeloma. Allogeneic hematopoietic stem cells and progenitors cells may be used, for example, for treating a recipient subject who has acute leukemia (e.g., AML, ALL); chronic lymphocytic leukemia (CLL); amegakaryocytosis/congenital thrombocytopenia; aplastic anemia/refractory anemia; familial erythrophagocytic lymphohistiocytosis; myelodysplastic syndrome/other myelodysplastic disorders; osteopetrosis; paroxysmal nocturnal hemoglobinuria; and Wiskott-Aldrich syndrome, for example. Exemplary uses for autologous hematopoietic stem cells and progenitors cells include treating a recipient subject who has amyloidosis; germ cell tumors (e.g., testicular cancer); or a solid tumor. Allogeneic hematopoietic stem cell transplants have also been investigated for use in treating solid tumors (see, e.g., Ueno et al., Blood 102:3829-36 (2003)).

In some embodiments of the methods described herein, the subject is not a donor of peripheral hematopoietic cells but has a disease, disorder, or condition for which mobilization of hematopoietic cells in the subject will provide clinical benefit. Stated another way, while this clinical situation is similar to autologous hematopoietic cell replacement, the mobilized hematopoietic cells are not removed and given back to the same subject at a later time as occurs, for example, with a subject who receives myeloablation therapy. Accordingly, methods are provided for mobilizing hematopoietic cells, such as hematopoietic stem cells and progenitor cells and leukocytes (including granulocytes, such as neutrophils), by administering at least once compound of Formula (I). Mobilizing hematopoietic stem cells and progenitor cells may be useful for treating an inflammatory condition or for tissue repair or wound healing. See, e.g., Mimeault et al., Clin. Pharmacol. Therapeutics 82:252-64 (2007).

In some embodiments, the methods described herein are useful for mobilizing hematopoietic leukocytes (white blood cells) in a subject, which methods may be used in treating diseases, disorders, and conditions for which an increase in white blood cells, such as neutrophils, eosinophils, lymphocytes, monocytes, basophils, will provide clinical benefit. For example, for cancer patients, the compounds of Formula (I) are beneficial for stimulating neutrophil production to compensate for hematopoietic deficits resulting from chemotherapy or radiation therapy. Other diseases, disorders, and conditions to be treated include infectious diseases and related conditions, such as sepsis. When the subject to whom at least one compound of Formula (I) is administered is a donor, neutrophils may be collected for administration to a recipient subject who has reduced hematopoietic function, reduced immune function, reduced neutrophil count, reduced neutrophil mobilization, severe chronic neutropenia, leucopenia, thrombocytopenia, anemia, and acquired immune deficiency syndrome. Mobilization of mature white blood cells may be useful in subjects to improve or to enhance tissue repair, and to minimize or prevent vascular injury and tissue damage, for example following liver transplantation, myocardial infarction or limb ischemia. See, e.g., Pelus, Curr. Opin. Hematol. 15:285-92 (2008); Lemoli et al., Haematologica 93: 321-24 (2008).

The compounds of Formula (I) may be used in combination with one or more other agents that mobilize hematopoietic cells. Such agents include, for example, G-CSF; AMD3100 or other CXCR4 antagonists; GRO-β (CXCL2) and an N-terminal 4-amino truncated form (SB-251353); IL-8SDF-1α peptide analogs, CTCE-0021 and CTCE-0214; and the SDF1 analog, Met-SDF-1β (see, e.g., Pelus, supra and references cited therein). In some embodiments, a compound of Formula (I) may be administered with other mobilizing agents used in the art, which may permit administration of a lower dose of GCSF or AMD3100, for example, than required in the absence of a compound of Formula (I). The appropriate therapeutic regimen for administering a compound of Formula (I) in combination with another mobilizing agent or agents can be readily determined by a person skilled in the clinical art.

The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may be useful in methods for preventing and/or treating mucositis. In some embodiments, at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) may be used in methods described herein for decreasing the likelihood of occurrence of mucositis in a subject who is in need thereof by administering the compound or composition to the subject. In some embodiments, the mucositis is chosen from oral mucositis, esophageal mucositis, and gastrointestinal mucositis. In some embodiments, the mucositis is alimentary mucositis.

It is believed that approximately half of all cancer patients undergoing therapy suffer some degree of mucositis. Mucositis is believed to occur, for example, in virtually all patients treated with radiation therapy for head and neck tumors, all patients receiving radiation along the GI tract, and approximately 40% of those subjected to radiation therapy and/or chemotherapy for tumors in other locations (e.g., leukemias or lymphomas). It is also is believed to be highly prevalent in patients treated with high dose chemotherapy and/or irradiation for the purpose of myeloablation, such as in preparation for stem cell or bone marrow transplantation. The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may be useful in methods for treating and/or preventing mucositis in a subject afflicted with cancer. In some embodiments, the subject is afflicted with a cancer chosen from head and neck cancer, breast cancer, lung cancer, ovarian cancer, prostate cancer, lymphatic cancer, leukemic cancer, and/or gastrointestinal cancer. In some embodiments, the mucositis is associated with radiation therapy and/or chemotherapy. In some embodiments, the chemotherapy comprises administering a therapeutically effective amount of at least one compound chosen from platinum, cisplatin, carboplatin, oxaliplatin, mechlorethamine, cyclophosphamide, chlorambucil, azathioprine, mercaptopurine, vincristine, vinblastine, vinorelbine, vindesine, etoposide, teniposide, paclitaxel, docetaxel, irinotecan, topotecan, amsacrine, etoposide, etoposide phosphate, teniposide, 5-fluorouracil (5-FU), leucovorin, methotrexate, gemcitabine, taxane, leucovorin, mitomycin C, tegafur-uracil, idarubicin, fludarabine, mitoxantrone, ifosfamide and doxorubicin.

In some embodiments, the method further comprises administering a therapeutically effective amount of at least one MMP inhibitor, inflammatory cytokine inhibitor, mast cell inhibitor, NSAID, NO inhibitor, or antimicrobial compound.

In some embodiments, the method further comprises administering a therapeutically effective amount of velafermin and/or palifermin.

In some embodiments, the method further comprises administering a therapeutically effective amount of Davanat®, mannose, and/or galactose.

The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may be useful in methods for treating and/or preventing thrombosis. As described herein methods are provided for inhibiting formation of a thrombus or inhibiting the rate at which a thrombus is formed. These methods may therefore be used for preventing thrombosis (i.e., reducing or decreasing the likelihood of occurrence of a thrombus in a statistically or clinically significant manner).

Thrombus formation may occur in infants, children, teenagers and adults. An individual may have a hereditary predisposition to thrombosis. Thrombosis may be initiated, for example, due to a medical condition (such as cancer or pregnancy), a medical procedure (such as surgery) or an environmental condition (such as prolonged immobility). Other individuals at risk for thrombus formation include those who have previously presented with a thrombus.

The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may be useful in methods for treating individuals undergoing thrombosis or who are at risk of a thrombotic event occurring. Such individuals may or may not have a risk of bleeding. In some embodiments, the individual has a risk of bleeding. In some embodiments, the thrombosis is a venous thromboembolism (VTE). VTE causes deep vein thrombosis and pulmonary embolism. Low molecular weight (LMW) heparin is the current mainstay therapy for the prevention and treatment of VTE. In many circumstances, however, the use of LMW heparin is contraindicated. LMW heparin is a known anti-coagulant and delays clotting over four times longer than control bleeding times. Patients undergoing surgery, patients with thrombocytopenia, patients with a history of stroke, and many cancer patients should avoid administration of heparin due to the risk of bleeding. By contrast, administration of the compounds of Formula (I) significantly reduces the time to clotting than occurs when LMW heparin is administered, and thus provide a significant improvement in reducing bleeding time compared with LMW heparin. Accordingly, the compounds and pharmaceutical compositions described herein may not only be useful for treating a patient for whom the risk of bleeding is not significant, but also may be useful in when the risk of bleeding is significant and the use of anti-thrombosis agents with anti-coagulant properties (such as LMW heparin) is contraindicated.

The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may be administered in combination with at least one additional anti-thrombosis agent. The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may function independently from the anti-thrombosis agent or may function in coordination with the at least one anti-thrombosis agent. In addition, the administration of one or more of the compounds or compositions may be in conjunction with one or more other therapies, e.g., for reducing toxicities of therapy. For example, at least one palliative agent to counteract (at least in part) a side effect of therapy may be administered. Agents (chemical or biological) that promote recovery and/or counteract side effects of administration of antibiotics or corticosteroids are examples of such palliative agents. The compounds of the present disclosure and pharmaceutical composition comprising at least one such compound may be administered before, after, or concurrently with administration of at least one additional anti-thrombosis agent or at least one palliative agent to reduce a side effect of therapy. Where administration is concurrent, the combination may be administered from a single container or two (or more) separate containers.

The compounds of the present disclosure and pharmaceutical compositions comprising at least one such compound may be useful for treating and/or preventing at least one cardiovascular disease, disorder and/or condition. Non-limiting examples of cardiovascular disease include atherosclerosis, myocardial infarction, myocardial ischemia, coronary artery stenosis (occlusion of the coronary arteries), chronic cardiovascular and/or arterial inflammation, acute cardiovascular and/or arterial inflammation, hypercholesterolemia, restenosis (narrowing of the vessel lumen), arrhythmia, thrombosis, hyperlipidemia, hypertension, dyslipoproteinemia, angina (cardiac chest pain), and vascular complications due to a cardiovascular disease (e.g., myocardial infarction or myocardial ischemia).

In some embodiments, at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) may be administered prior to or subsequent to an acute cardiovascular event in the subject. In some embodiments, at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) may be administered prior to or subsequent to the development or diagnosis of a cardiovascular disease, disorder and/or condition in the subject. In some embodiments, the acute cardiovascular event is a myocardial infarction.

In some embodiments, a method for treatment and/or prevention of atherosclerosis is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I). Atherosclerosis generally describes a disease of the arterial blood vessels. As used herein, “atherosclerosis” includes, but is not limited to, chronic and/or acute atherosclerotic inflammation prior to or subsequent to the formation of at least one atherosclerotic plaque in the subject. Atherosclerosis also includes, but is not limited to, chronic progressive atherosclerosis and/or atherosclerotic inflammation. Atherosclerosis also includes, but is not limited to, acute atherosclerosis and/or atherosclerotic inflammation subsequent to an acute vascular event in the subject (such as, for example, myocardial infarction).

In some embodiments, at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) may be administered prior to or subsequent to the formation of at least one atherosclerotic plaque, lesion or atheroma in the subject.

In some embodiments, the formation, progression, destabilization and/or rupture of at least one atherosclerotic plaque within the subject is reduced.

Atherosclerotic plaques may be characterized as stable or unstable (i.e., vulnerable to destabilization). Unstable atherosclerotic plaques may be susceptible to disruption or rupture, which exposes thrombogenic material (i.e., thrombi) (e.g., collagen) to the circulation. This can produce interruptions in blood flood (ischemia) in local or distal arteries, which can result in cardiovascular complications, such as, for example, myocardial infarction (MI).

Destabilization of atherosclerotic plaques may occur via many mechanisms. Non-limiting examples of such mechanisms include superficial erosion of the endothelial cells that form the monolayer covering the intima, disruption of the microvessels that form in the atherosclerotic plaque, rupture (i.e., fracture) of the atherosclerotic plaque's fibrous cap, thinning or weakening of the fibrous cap (thus making it susceptible to rupture), and the presence or increase in inflammatory factors that mediate destabilization. (Libby P., Nature, 420: 868-874 (2002)).

A non-limiting example of inflammatory factors that mediate destabilization is the presence of inflammatory cells. The progression of atherosclerosis may be associated with systemically increased inflammatory myeloid cells that are recruited to atherosclerotic plaques. (Murphy, A. J. et al., J. Clin. Invest., 121: 4138-4149 (2011); Averill, L. E. et al., Am. J. Pathol., 135: 369-377 (1989); Feldman, D. L. et al., Arterioscler. Thromb., 11: 985-994 (1991); Swirski, F. K. et al., J. Clin. Invest. 117: 195-205 (2007)). The presence of inflammatory myeloid cells may be detrimental to a stable plaque. (Llodra, J. et al., Proc. Natl. Acad. Sci. U.S.A., 101: 11779-11784 (2004)).

In some embodiments, the stability of at least one atherosclerotic plaque within the subject is increased. Non-limiting examples of stable features of atherosclerotic plaques (i.e., stable phenotype) include smaller plaque size, reduced (i.e., decreased, diminished, smaller) necrotic core size (measured by, for example, necrotic core area), and a thicker fibrous cap of the atherosclerotic plaque. (See, e.g., Moore K. J. et al., Cell, 145: 341-355 (2011)).

In some embodiments, the size of at least one atherosclerotic plaque within the subject is decreased. In some embodiments, the necrotic core size of at least one atherosclerotic plaque within the subject is decreased. In some embodiments, the fibrous cap thickness of at least one atherosclerotic plaque within the subject is increased.

In some embodiments, the administration of an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) reduces the levels of extramedullary proliferation of hematopoietic stem and/or progenitor cells within the subject. In some embodiments, extramedullary proliferation of hematopoietic stem and/or progenitor cells is reduced in the spleen and/or the liver. Non-limiting examples of extramedullary proliferation of hematopoietic stem and/or progenitor cells include extramedullary hematopoiesis and extramedullary myelopoiesis.

In some embodiments, the administration of an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) reduces the recruitment and/or infiltration of myeloid cells to at least one atherosclerotic plaque within the subject. Non-limiting examples of myeloid cells include monocytes, macrophages, neutrophils, basophils, eosinophils, erythrocytes, dendritic cells, and megakaryocytes and platelets.

In some embodiments, the at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) is administered after angioplasty, stenting procedure, atherectomy, bypass surgery, or other vessel-corrective techniques.

In some embodiments, the at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) is administered before angioplasty, stenting procedure, atherectomy, bypass surgery, or other vessel-corrective techniques.

In some embodiments, a method for treatment and prevention of myocardial infarction is disclosed, the method comprising administering to a subject in need thereof an effective amount of at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I). In some embodiments, the subject has previously suffered a myocardial infarction. In some embodiments, a compound of Formula (I) may be administered before the occurrence of a myocardial infarction in the subject. In some embodiments, a compound of Formula (I) may be administered after the occurrence of a first or subsequent myocardial infarction in the subject.

In some embodiments, at least one compound of Formula (I) and/or a pharmaceutical composition comprising at least one compound of Formula (I) is administered to the subject: within one (1) day of the subject suffering a myocardial infarction, within one (1) week of the subject suffering a myocardial infarction, within two (2) weeks of the subject suffering a myocardial infarction, within three (3) weeks of the subject suffering a myocardial infarction, within four (4) weeks of the subject suffering a myocardial infarction, within eight (8) weeks of the subject suffering a myocardial infarction, or within twelve (12) weeks of the subject suffering a myocardial infarction.

The terms, “treat” and “treatment,” include medical management of a disease, disorder, and/or condition of a subject as would be understood by a person of ordinary skill in the art (see, e.g., Stedman's Medical Dictionary). In general, an appropriate dose and treatment regimen provide at least one of the compounds of the present disclosure in an amount sufficient to provide therapeutic and/or prophylactic benefit. For both therapeutic treatment and prophylactic or preventative measures, therapeutic and/or prophylactic benefit includes, for example, an improved clinical outcome, wherein the object is to prevent or slow or lessen an undesired physiological change or disorder, or to prevent or slow or lessen the expansion or severity of such disorder. As discussed herein, beneficial or desired clinical results from treating a subject include, but are not limited to, abatement, lessening, or alleviation of symptoms that result from or are associated with the disease, condition, and/or disorder to be treated; decreased occurrence of symptoms; improved quality of life; longer disease-free status (i.e., decreasing the likelihood or the propensity that a subject will present symptoms on the basis of which a diagnosis of a disease is made); diminishment of extent of disease; stabilized (i.e., not worsening) state of disease; delay or slowing of disease progression; amelioration or palliation of the disease state; and remission (whether partial or total), whether detectable or undetectable; and/or overall survival. “Treatment” can include prolonging survival when compared to expected survival if a subject were not receiving treatment.

In some embodiments of the methods described herein, the subject is a human. In some embodiments of the methods described herein, the subject is a non-human animal. Non-human animals that may be treated include mammals, for example, non-human primates (e.g., monkey, chimpanzee, gorilla, and the like), rodents (e.g., rats, mice, gerbils, hamsters, ferrets, rabbits), lagomorphs, swine (e.g., pig, miniature pig), equine, canine, feline, bovine, and other domestic, farm, and zoo animals.

The effectiveness of the compounds of the present disclosure in treating and/or preventing diseases, disorders, and/or conditions treatable by inhibiting an activity of E-selectin can readily be determined by a person of ordinary skill in the relevant art. Determining and adjusting an appropriate dosing regimen (e.g., adjusting the amount of compound per dose and/or number of doses and frequency of dosing) can also readily be performed by a person of ordinary skill in the relevant art. One or any combination of diagnostic methods, including physical examination, assessment and monitoring of clinical symptoms, and performance of analytical tests and methods described herein, may be used for monitoring the health status of the subject.

Also provided herein are pharmaceutical compositions comprising at least one compound of Formula (I). In some embodiments, the pharmaceutical compositions further comprise at least one additional pharmaceutically acceptable ingredient.

In pharmaceutical compositions, any one or more of the compounds of the present disclosure may be administered in the form of a pharmaceutically acceptable derivative, such as a salt, and/or it or they may also be used alone and/or in appropriate association, as well as in combination, with other pharmaceutically active compounds.

An effective amount or therapeutically effective amount refers to an amount of at least one compound of the present disclosure or a pharmaceutical composition comprising at least one such compound that, when administered to a subject, either as a single dose or as part of a series of doses, is effective to produce at least one therapeutic effect. Optimal doses may generally be determined using experimental models and/or clinical trials. Design and execution of pre-clinical and clinical studies for each of the therapeutics (including when administered for prophylactic benefit) described herein are well within the skill of a person of ordinary skill in the relevant art. The optimal dose of a therapeutic may depend upon the body mass, weight, and/or blood volume of the subject. In general, the amount of at least one compound of Formula (I) as described herein, that is present in a dose, may range from about 0.1 mg to about 100 mg per kg weight of the subject. The minimum dose that is sufficient to provide effective therapy may be used in some embodiments. Subjects may generally be monitored for therapeutic effectiveness using assays suitable for the disease, disorder and/or condition being treated or prevented, which assays will be familiar to those having ordinary skill in the art and are described herein. The level of a compound that is administered to a subject may be monitored by determining the level of the compound (or a metabolite of the compound) in a biological fluid, for example, in the blood, blood fraction (e.g., serum), and/or in the urine, and/or other biological sample from the subject. Any method practiced in the art to detect the compound, or metabolite thereof, may be used to measure the level of the compound during the course of a therapeutic regimen.

The dose of a compound described herein may depend upon the subject's condition, that is, stage of the disease, severity of symptoms caused by the disease, general health status, as well as age, gender, and weight, and other factors apparent to a person of ordinary skill in the medical art. Similarly, the dose of the therapeutic for treating a disease, disorder, and/or condition may be determined according to parameters understood by a person of ordinary skill in the medical art.

Pharmaceutical compositions may be administered in any manner appropriate to the disease, disorder, and/or condition to be treated as determined by persons of ordinary skill in the medical arts. An appropriate dose and a suitable duration and frequency of administration will be determined by such factors as discussed herein, including the condition of the patient, the type and severity of the patient's disease, the particular form of the active ingredient, and the method of administration. In general, an appropriate dose (or effective dose) and treatment regimen provides the composition(s) as described herein in an amount sufficient to provide therapeutic and/or prophylactic benefit (for example, an improved clinical outcome, such as more frequent complete or partial remissions, or longer disease-free and/or overall survival, or a lessening of symptom severity or other benefit as described in detail above).

The pharmaceutical compositions described herein may be administered to a subject in need thereof by any one of several routes that effectively delivers an effective amount of the compound. Non-limiting examples of suitable administrative routes include topical, oral, nasal, intrathecal, enteral, buccal, sublingual, transdermal, rectal, vaginal, intraocular, subconjunctival, sublingual, and parenteral administration, including subcutaneous, intravenous, intramuscular, intrasternal, intracavernous, intrameatal, and intraurethral injection and/or infusion.

The pharmaceutical compositions described herein may, for example, be sterile aqueous or sterile non-aqueous solutions, suspensions, or emulsions, and may additionally comprise at least one pharmaceutically acceptable excipient (i.e., a non-toxic material that does not interfere with the activity of the active ingredient). Such compositions may, for example, be in the form of a solid, liquid, or gas (aerosol). Alternatively, the compositions described herein may, for example, be formulated as a lyophilizate, or compounds described herein may be encapsulated within liposomes using technology known in the art. The pharmaceutical compositions may further comprise at least one additional pharmaceutically acceptable ingredient, which may be biologically active or inactive. Non-limiting examples of such ingredients include buffers (e.g., neutral buffered saline or phosphate buffered saline), carbohydrates (e.g., glucose, mannose, sucrose or dextrans), mannitol, proteins, polypeptides, amino acids (e.g., glycine), antioxidants, chelating agents (e.g., EDTA and glutathione), stabilizers, dyes, flavoring agents, suspending agents, and preservatives.

Any suitable excipient or carrier known to those of ordinary skill in the art for use in compositions may be employed in the compositions described herein. Excipients for therapeutic use are well known, and are described, for example, in Remington: The Science and Practice of Pharmacy (Gennaro, 21^(st) Ed. Mack Pub. Co., Easton, Pa. (2005)). In general, the type of excipient may be selected based on the mode of administration, as well as the chemical composition of the active ingredient(s). Compositions may be formulated for the particular mode of administration. For parenteral administration, pharmaceutical compositions may further comprise water, saline, alcohols, fats, waxes, and buffers. For oral administration, pharmaceutical compositions may further comprise at least one component chosen, for example, from any of the aforementioned ingredients, excipients and carriers, such as mannitol, lactose, starch, magnesium stearate, sodium saccharine, talcum, cellulose, kaolin, glycerin, starch dextrins, sodium alginate, carboxymethylcellulose, ethyl cellulose, glucose, sucrose, and magnesium carbonate.

The pharmaceutical compositions (e.g., for oral administration or delivery by injection) may be in the form of a liquid. A liquid composition may include, for example, at least one the following: a sterile diluent such as water for injection, saline solution, including for example physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils that may serve as the solvent or suspending medium, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents; antioxidants; chelating agents; buffers and agents for the adjustment of tonicity such as sodium chloride or dextrose. A parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. In some embodiments, the pharmaceutical composition comprises physiological saline. In some embodiments, the pharmaceutical composition is an injectable composition, and in some embodiments, the injectable composition is sterile.

For oral formulations, at least one of the compounds of the present disclosure can be used alone or in combination with at least one additive appropriate to make tablets, powders, granules and/or capsules, for example, those chosen from conventional additives, disintegrators, lubricants, diluents, buffering agents, moistening agents, preservatives, coloring agents, and flavoring agents. The pharmaceutical compositions may be formulated to include at least one buffering agent, which may provide for protection of the active ingredient from low pH of the gastric environment and/or an enteric coating. A pharmaceutical composition may be formulated for oral delivery with at least one flavoring agent, e.g., in a liquid, solid or semi-solid formulation and/or with an enteric coating.

Oral formulations may be provided as gelatin capsules, which may contain the active compound or biological along with powdered carriers. Similar carriers and diluents may be used to make compressed tablets. Tablets and capsules can be manufactured as sustained release products to provide for continuous release of active ingredients over a period of time. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.

A pharmaceutical composition may be formulated for sustained or slow release. Such compositions may generally be prepared using well known technology and administered by, for example, oral, rectal or subcutaneous implantation, or by implantation at the desired target site. Sustained-release formulations may contain the active therapeutic dispersed in a carrier matrix and/or contained within a reservoir surrounded by a rate controlling membrane. Excipients for use within such formulations are biocompatible, and may also be biodegradable; the formulation may provide a relatively constant level of active component release. The amount of active therapeutic contained within a sustained release formulation depends upon the site of implantation, the rate and expected duration of release, and the nature of the condition to be treated or prevented.

The pharmaceutical compositions described herein can be formulated as suppositories by mixing with a variety of bases such as emulsifying bases or water-soluble bases. The pharmaceutical compositions may be prepared as aerosol formulations to be administered via inhalation. The pharmaceutical compositions may be formulated into pressurized acceptable propellants such as dichlorodifluoromethane, propane, nitrogen and the like.

The compounds of the present disclosure and pharmaceutical compositions comprising these compounds may be administered topically (e.g., by transdermal administration). Topical formulations may be in the form of a transdermal patch, ointment, paste, lotion, cream, gel, and the like. Topical formulations may include one or more of a penetrating agent or enhancer (also call permeation enhancer), thickener, diluent, emulsifier, dispersing aid, or binder. Physical penetration enhancers include, for example, electrophoretic techniques such as iontophoresis, use of ultrasound (or “phonophoresis”), and the like. Chemical penetration enhancers are agents administered either prior to, with, or immediately following administration of the therapeutic, which increase the permeability of the skin, particularly the stratum corneum, to provide for enhanced penetration of the drug through the skin. Additional chemical and physical penetration enhancers are described in, for example, Transdermal Delivery of Drugs, A. F. Kydonieus (ED) 1987 CRL Press; Percutaneous Penetration Enhancers, eds. Smith et al. (CRC Press, 1995); Lenneräs et al., J. Pharm. Pharmacol. 54:499-508 (2002); Karande et al., Pharm. Res. 19:655-60 (2002); Vaddi et al., Int. J. Pharm. 91:1639-51 (2002); Ventura et al., J. Drug Target 9:379-93 (2001); Shokri et al., Int. J. Pharm. 228(1-2):99-107 (2001); Suzuki et al., Biol. Pharm. Bull. 24:698-700 (2001); Alberti et al., J. Control Release 71:319-27 (2001); Goldstein et al., Urology 57:301-5 (2001); Kiijavainen et al., Eur. J. Pharm. Sci. 10:97-102 (2000); and Tenjarla et al., Int. J. Pharm. 192:147-58 (1999).

Kits comprising unit doses of at least one compound of the present disclosure, for example in oral or injectable doses, are provided. Such kits may include a container comprising the unit dose, an informational package insert describing the use and attendant benefits of the therapeutic in treating the pathological condition of interest, and/or optionally an appliance or device for delivery of the at least one compound of Formula (I) and/or pharmaceutical composition comprising the same.

EXAMPLES

Unless otherwise noted, all chemicals were reagent grade materials from commercial suppliers and were used without further purification. NMR data were recorded on a Bruker 300 MHz spectrometer or a Bruker 400 MHz spectrometer and the chemical shifts are reported in ppm. Coupling constants (J) are reported in hertz. Mass spectra were recorded on an Agilent 1100 Series LC/MSD spectrometer.

Example 1 Synthesis of Compound 17

Compound 3: A solution of compound 3 was prepared according to Bioconjugate. Chem. 2014, 25, 1444-52.

Compound 4: Sulfur trioxide pyridine complex was purified by placing the material in a fritted funnel and washing with ice water until the pH of the filtrate was between 5-6. The material was then dried by washing with ice cold ethanol followed by dichloromethane and then ether. The material was used immediately for reaction.

3-Azidopropanol (0.56 g, 5.6 mmol) was dissolved in dry pyridine (20 mL) under an argon atmosphere. Freshly purified sulfur trioxide pyridine complex (2.92 g, 18.3 mmol) was added and the reaction mixture was stirred overnight at 60° C. The solvent was removed and the resulting solid was dissolved in 10 mL de-ionized water. The solution was cooled on an ice bath and the pH was adjusted to ˜9 by slow addition of 3N NaOH. The solvent was removed and the resulting solid was suspended in isopropanol and filtered to afford 410 mg compound 4 as a yellow-brown solid. MS negative mode: m/z=180.0 [M-Na]⁻ C₃H₆N₃NaO₄S (203).

¹H NMR (400 MHz, D₂O) δ 4.04 (t, J=6.0 Hz, 2H), 3.37 (t, J=6.6 Hz, 2H), 1.86 (p, J=6.3 Hz, 2H).

Compound 6: Prepared according to Bioorg. Med. Chem. Lett. 1995, 5, 2321-2324 starting with D-threonolactone.

Compound 7: Compound 6 (500 mg, 1 mmol) was dissolved in 9 mL acetonitrile. Potassium hydroxide (1 mL of a 2M solution) was added and the reaction mixture was stirred at 50° C. for 12 hours. The reaction mixture was partitioned between dichloromethane and water. The phases were separated and the aqueous phase was extracted 3 times with dichloromethane. The aqueous phase was acidified with 1N HCl until pH˜1 and extracted 3 times with dichloromethane. The combined dichloromethane extracts from after acidification of the aqueous phase were concentrated in vacuo to give compound 7 as a yellow oil (406 mg). LCMS (C-18; 5-95 H₂O/MeCN): UV (peak at 4.973 min), positive mode: m/z=407 [M+H]⁺; negative mode: m/z=405 [M−H]⁻ C₂₅H₂₆O₅ (406).

Compound 11: Compound 47 (prepared according to WO 2013/096926) (0.18 g) was dissolved in ethylenediamine (2 mL) and stirred at 80° C. for 8 hours. The solvent was removed in vacuo and the crude material separated by C-18 reverse phase chromatography to afford 0.15 g of compound 11. MS positive mode: m/z=756.3 [M+H]⁺. MS negative mode: m/z=732.3 [M-Na]⁻ C₃₄H₅₈N₃NaO₁₄ (755.38).

¹H NMR (400 MHz, D₂O) δ 5.02 (d, J=3.9 Hz, 1H), 4.89 (q, J=6.6 Hz, 1H), 4.54 (d, J=8.5 Hz, 1H), 3.95 (t, J=8.1 Hz, 1H), 3.89-3.85 (m, 3H), 3.83-3.74 (m, 2H), 3.72 (d, J=5.8 Hz, 2H), 3.55 (t, J=5.9 Hz, 1H), 3.50-3.44 (m, 3H), 3.33 (t, J=9.7 Hz, 1H), 3.11 (t, J=6.1 Hz, 2H), 2.36 (t, J=12.6 Hz, 1H), 2.17 (br d, J=12.0 Hz, 1H), 2.04 (s, 3H), 1.90 (d, J=13.7 Hz, 1H), 1.87-1.83 (m, 1H), 1.76-1.66 (m, 2H), 1.64-1.41 (m, 3H), 1.41-1.28 (m, 1H), 1.22 (d, J=6.4 Hz, 3H), 1.20-1.14 (m, 3H), 0.92-0.87 (m, 2H), 0.84 (t, J=7.3 Hz, 3H).

Compound 12: To a stirred and cooled (15° C.) solution of propiolic acid (82 μl, 1.19 mmol) in DCM (5 mL) under argon was added oxalyl chloride (100 μl, 1.19 mmol) and DMF (5 μl, cat). The resulting mixture is stirred for 20 min before addition to a solution of compound 11 (200 mg, 0.27 mmol) in MeOH (20 mL) containing saturated sodium bicarbonate solution (4 mL). The resulting mixture was stirred at room temperature for 30 min; LCMS at that time showed the reaction was complete. The reaction was filtered and partially concentrated under reduced pressure to the water fraction, which, was in-turn freeze-dried. Gel permeation of the residue (water as eluent) afforded compound 12 as a white solid (214 mg) (100%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 3.342 min), UV (peak at 3.247 min), negative mode: m/z=785.2 [M−H]⁻ C₃₇H₅₉N₃O₁₅ (786).

¹H NMR (300 MHz, D₂O) δ 4.96 (d, J=3.9 Hz, 1H), 4.83 (q, J=8.0 Hz, 1H), 4.45 (d, J=8.6 Hz, 1H), 3.93 (t, J=9.5 Hz, 1H), 3.84-3.78 (m, 3H), 3.78-3.72 (m, 2H), 3.72-3.62 (m, 3H), 3.49 (t, J=5.9 Hz, 1H), 3.42-3.32 (m, 2H), 3.32-3.21 (m, 4H), 2.25 (br t, J=12.1 Hz, 1H), 2.09 (br d, J=7.3 Hz, 1H), 2.00 (s, 3H), 1.80 (br d, J=10.3 Hz, 2H), 1.74-1.37 (m, 7H), 1.37-1.20 (m, 3H), 1.17 (d, J=6.5 Hz, 3H), 1.14-1.01 (m, 3H), 0.93-0.83 (m, 2H), 0.79 (t, J=7.3 Hz, 3H).

Compound 17: A stirred solution of compound 12 (75 mg, 95 μmol) and azide 3 (288 mg, 114 μmol) in HPLC grade water (6 ml) under argon was treated with a pre-mixed solution of copper sulfate and THPTA (19 μmol, 0.5 mL of 40 mmol aqueous solution) and sodium ascorbate (124 μmol, 0.125 mL of 1M solution). After 2 h the reaction was lyophilized. Gel permeation of the residue (water as eluent) afforded compound 17 as an off-white solid (46 mg, 39%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 3.046 min), UV (peak at 2.846/2.952 min), negative mode: m/z=592 [(M-4Na)/2]⁻ C₄₂H₆₆N₆Na₃O₂₇S₃ (1274.27).

¹H NMR (300 MHz, D₂O) δ 8.47 (s, 1H), 4.95 (d, J=4.0 Hz, 1H), 4.85 (q, J=7.0 Hz, 1H), 4.43 (d, J=8.5 Hz, 1H), 4.15 (br s, 1H), 4.01 (s, 6H), 3.93 (br s, 1H), 3.81 (dd, J=10.5, 3.2 Hz, 1H), 3.74-3.69 (m, 2H), 3.69-3.57 (m, 3H), 3.54-3.41 (m, 1H), 3.41-3.30 (m, 1H), 3.24 (t, J=9.6 Hz, 1H), δ 2.22 (br t, J=12.4 Hz, 1H), 2.06 (br s, 1H), 1.99 (s, 3H), 1.84-1.61 (m, 2H), 1.60-1.39 (m, 7H), 1.26 (q, J=12.7, 12.2 Hz, 1H), 1.16 (d, J=6.5 Hz, 3H), 0.96 (dd, J=68.9, 12.0 Hz, 1H), 0.74 (t, J=7.0 Hz, 3H).

Example 2 Synthesis of Compound 18

Compound 13: Compound 11 (501 mg, 0.66 mmol, prepared according to WO 2013/096926) and triethylamine (0.18 mL, 1.32 mmol) were dissolved in 5 mL MeOH and cooled on an ice bath. Ethyl trifluoroacetate (0.1 mL, 0.79 mmol) was added dropwise and the reaction mixture was stirred 1 hour on the ice bath. The solvent was removed and the residue was azeotroped three times from toluene to afford compound 13. The crude material was used without further purification. MS m/z=852.3 [M+H]: C₃₆H₅₇F₃N₃NaO₁₅.

¹H NMR (400 MHz, DMSO-d6) δ 9.41 (t, J=5.4 Hz, 1H), 7.94 (t, J=5.4 Hz, 1H), 7.63 (br s, 1H), 4.76 (q, J=6.5 Hz, 1H), 4.67 (d, J=3.5 Hz, 1H), 4.44 (br s, 2H), 4.16 (d, J=6.5 Hz, 2H), 4.11 (d, J=4.4 Hz, 1H), 3.68 (br s, 1H), 3.64 (d, J=8.7 Hz, 1H), 3.58-3.47 (m, 3H), 3.45-3.37 (m, 2H), 3.28-3.19 (m, 3H), 2.89 (br s, 4H), 2.11 (br t, I=12.05 Hz, 1H), 2.04 (br d, J=12.3 Hz, 1H), 1.99-1.90 (m, 1H), 1.77 (s, 3H), 1.72-1.56 (m, 2H), 1.52 (br d, J=13.1 Hz, 1H), 1.46-1.36 (m, 3H), 1.20 (q, J=12.1 Hz, 2H), 1.06 (d, J=6.3 Hz, 3H), 1.04-0.84 (m, 2H), 0.79 (t, J=7.4 Hz, 3H).

Compound 14: Crude compound 13 (0.66 mmol) was dissolved in dry DMF and cooled on an ice bath under an argon atmosphere. Diisopropylethylamine (0.23 mL, 1.32 mmol) was added followed by HATU (300 mg, 0.79 mmol). The reaction mixture was stirred 15 minutes. Azetidine (90 μL, 1.32 mmol) was added and the reaction mixture was stirred overnight allowing to warm to room temperature during which time the product precipitated from the solution. The reaction mixture was suspended in acetonitrile, stirred 1 minute and the mixture allowed to settle. The yellow acetonitrile solution was carefully removed using a pipet. This procedure was repeated until the acetonitrile solution was no longer yellow then 2 more times. The remaining solid was dried in vacuo to afford 502 mg compound 14 (87% yield for 2 steps). MS m/z=891.3 [M+Na]: C₃₉H₆₃F₃N₄NaO₁₄.

¹H NMR (400 MHz, DMSO-d6) δ 9.40 (t, J=2.5 Hz, 1H), 7.93 (t, J=6.1 Hz, 1H), 7.70 (br s, 1H), 4.75 (q, J=6.7 Hz, 1H), 4.67 (d, J=3.3 Hz, 1H), 4.55 (t, J=5.5 Hz, 1H), 4.31 (d, J=2.9 Hz, 1H), 4.26 (q, J=7.9 Hz, 1H), 4.19-4.14 (m, 2H), 4.13-4.09 (m, 2H), 3.94 (br d, J=9.5 Hz, 1H), 3.87 (q, J=7.5 Hz, 1H), 3.63 (s, 1H), 3.56-3.49 (m, 2H), 3.46-3.36 (m, 3H), 3.29-3.18 (m, 3H), 3.14-3.03 (m, 2H), 2.22 (p, J=7.7 Hz, 2H), 2.11 (br t, J=12.1 Hz, 0H), 2.02 (br d, J=13.1 Hz, 1H), 1.98-1.90 (m, 1H), 1.79 (s, 3H), 1.74-1.50 (m, 4H), 1.49-1.34 (m, 1H), 1.29-1.08 (m, 2H), 1.05 (d, J=6.4 Hz, 3H), 1.02-0.84 (m, 2H), 0.79 (t, J=7.4 Hz, 3H).

Compound 15: Compound 14 (498 mg, 0.57 mmol) was suspended in 5 mL MeOH and cooled on an ice bath. Sodium methoxide (0.15 mL of a 25 wt % in MeOH solution, 0.68 mmol) was added and the reaction mixture was stirred allowing to warm to room temperature. The reaction mixture slowly became homogeneous. After stirring overnight at room temperature, MS indicates compound 13 is still present. Sodium methoxide solution (50 μL) was added and the reaction mixture was stirred at room temperature for another 5 hours. The reaction mixture was quenched by addition of 0.5 mL HOAc. The volatiles were removed in vacuo and the residue was separated by reverse phase chromatography using a 10 g C-18 cartridge and eluting with water then 3/1 water/MeOH then 1/1 water/MeOH. The product containing fractions were concentrated and the residue lyophilized to afford 420 mg of compound 15 as an off-white solid (94% yield). MS m/z=773.4 [M+H]: C₃₇H₆₄N₄O₁₃.

¹H NMR (400 MHz, DMSO-d6) δ 7.91 (t, I=5.4 Hz, 1H), 4.67 (q, J=6.5 Hz, 1H), 4.59 (d, J=3.4 Hz, 1H), 4.25 (s, 1H), 4.20 (q, J=7.9 Hz, 1H), 4.04 (q, J=8.0 Hz, 1H), 3.86 (dd, J=10.2, 2.6 Hz, 1H), 3.81 (q, J=8.4 Hz, 1H), 3.56 (s, 1H), 3.51-3.41 (m, 3H), 3.40-3.28 (m, 5H), 3.19-3.11 (m, 2H), 3.10 (s, 3H), 2.99 (dd, J=11.2, 7.8 Hz, 1H), 2.68 (t, J=6.5 Hz, 2H), 2.43 (p, J=1.8 Hz, 3H), 2.15 (q, J=7.7 Hz, 1H), 2.11-2.01 (m, OH), 2.01-1.83 (m, 1H), 1.71 (s, 3H), 1.68-1.50 (m, 2H), 1.50-1.26 (m, 2H), 1.20-1.02 (m, 3H), 1.00-0.95 (m, 1H), 0.94-0.76 (m, OH), 0.72 (t, J=7.2 Hz, 1H).

Compound 16: Propiolic acid (200 μl, 3.23 mmol) was dissolved in DCM (5 mL) under argon and cooled on an ice bath. Oxalyl chloride (1.6 ml of a 2M solution in DCM, 3.2 mmol) and DMF (1 drop, cat) were added and the reaction mixture was stirred for 1 hour. Compound 15 (210 mg, 0.27 mmol) was dissolved in 5 mL of saturated aqueous sodium bicarbonate solution and cooled on an ice bath. The solution of propiolyl chloride was added and the reaction mixture was stirred 3 hours on an ice bath. Volatiles were removed in vacuo and the resulting aqueous reaction mixture was separated by reverse phase chromatography using a 10 g C-18 cartridge and eluting with water followed by 1/1 water/MeOH followed by 1/3 water/MeOH to give recovered compound 15 (in the 1/1 water/MeOH eluent) and partially purified product (in the 1/3 water/MeOH eluent). The partially purified product was further purified using a 5 g C-18 cartridge and eluting with water followed by 1/1 water/MeOH followed by 1/3 water/MeOH to afford 29 mg compound 16 as a white solid (13%). MS, positive mode: m/z=847.4 [M+Na] C₄₀H₆₄N₄O₁₄ (824).

¹H NMR (400 MHz, D₂O) δ 4.93 (d, J=4.0 Hz, 1H), 4.79 (q, J=6.7 Hz, 1H), 4.42 (d, J=8.6 Hz, 1H), 4.24 (q, J=8.7 Hz, 1H), 4.15 (q, J=8.7 Hz, 1H), 4.01 (dd, J=9.9, 2.4 Hz, 1H), 3.94 (q, J=8.2 Hz, 1H), 3.87 (d, J=11.3 Hz, 0H), 3.77 (dd, J=10.7, 3.3 Hz, 1H), 3.74-3.65 (m, 3H), 3.65-3.56 (m, 2H), 3.49-3.37 (m, 1H), 3.36-3.28 (m, 1H), 3.28-3.16 (m, 3H), 2.29-2.16 (m, 3H), 2.03 (d, J=8.0 Hz, 1H), 1.95 (s, 3H), 1.76 (d, J=13.3 Hz, 2H), 1.61 (q, J=12.9 Hz, 1H), 1.54-1.38 (m, 2H), 1.33-1.16 (m, 2H), 1.12 (d, J=6.5 Hz, 3H), 1.10-1.01 (m, 1H), 0.85 (q, J=10.4, 9.8 Hz, 1H), 0.76 (t, J=7.3 Hz, 3H).

Compound 18: Using an analogous procedure to the procedure in FIG. 5, compound 18 was prepared from compound 16 and azide 3. MS negative mode: m/z=407.6 [(M-3Na)/3]⁻ C₄₅H₇₂N₇Na₃O₂₆S₃ (1291).

¹H NMR (400 MHz, D₂O) δ 8.36 (s, 1H), 4.86 (d, J=4.0 Hz, 1H), 4.77 (q, J=6.7 Hz, 1H), 4.59 (s, 1H), 4.35 (d, J=8.2 Hz, 1H), 4.28 (q, J=8.7 Hz, 1H), 4.09 (q, J=8.7 Hz, 1H), 4.02-3.98 (m, 1H), 3.98-3.81 (m, 6H), 3.72 (dd, J=10.5, 3.3 Hz, 1H), 3.68 (d, J=2.7 Hz, 1H), 3.66-3.58 (m, 2H), 3.58-3.49 (m, 3H), 3.46-3.35 (m, 2H), 3.26-3.13 (m, 2H), 3.08 (br t, J=11.6 Hz, 1H), 2.28-2.10 (m, 1H), 2.05-1.94 (m, 1H), 1.93 (s, 3H), 1.75-1.55 (m, 2H), 1.54-1.29 (m, 3H), 1.26-1.10 (m, 2H), 1.07 (d, J=6.5 Hz, 3H), 1.05-0.70 (m, 2H), 0.67 (t, J=7.3 Hz, 3H).

Example 3 Synthesis of Compound 19

Compound 4: Sulfur trioxide pyridine complex was purified by placing the material in a fritted funnel and washing with ice water until the pH of the filtrate was between 5-6. The material was then dried by washing with ice cold ethanol followed by dichloromethane and then ether. The material was used immediately for reaction.

3-Azidopropanol (0.56 g, 5.6 mmol) was dissolved in dry pyridine (20 mL) under an argon atmosphere. Freshly purified sulfur trioxide pyridine complex (2.92 g, 18.3 mmol) was added and the reaction mixture was stirred overnight at 60° C. The solvent was removed and the resulting solid was dissolved in 10 mL de-ionized water. The solution was cooled on an ice bath and the pH was adjusted to ˜9 by slow addition of 3N NaOH. The solvent was removed and the resulting solid was suspended in isopropanol and filtered to afford 410 mg compound 4 as a yellow-brown solid. MS negative mode: m/z=180.0 [M-Na]⁻ C₃H₆N₃NaO₄S (203).

¹H NMR (400 MHz, D₂O) δ 4.04 (t, J=6.0 Hz, 2H), 3.37 (t, J=6.6 Hz, 2H), 1.86 (p, J=6.3 Hz, 2H).

Compound 19: Compound 19 was prepared in a procedure analogous to the procedure in FIG. 5, using compound 16 and compound 4 as starting materials. MS negative mode: m/z=1004.4 [(M-Na)]⁻ C₄₃H₇₀N₇NaO₁₈S (1027).

¹H NMR (400 MHz, D₂O) δ 8.36 (s, 1H), 4.95 (d, J=4.0 Hz, 1H), 4.83 (q, J=8.0 Hz, 1H), 4.57 (t, J=8.0 Hz, 2H), 4.44 (broad d, J=8.0 Hz, 1H), 4.31 (q, J=8.0 Hz, 1H), 4.19 (q, J=8.0 Hz, 1H), 4.11-4.04 (m, 1H), 4.03-3.86 (m, 5H), 3.80 (dd, J=12.0, 4.0 Hz, 1H), 3.76 (broad d, 1H), 3.72 (broad d, 2H), 3.69-3.54 (m, 4H), 3.53-3.45 (m, 2H), 3.43-3.32 (m, 3H), 3.24 (br t, J=8.0 Hz, 1H), 2.36-2.17 (m, 5H), 2.12-2.03 (m, 1H), 1.97 (s, 3H), 1.83-1.40 (m, 9H), 1.37-0.97 (m, J=8.0 Hz, 11H), 0.96-0.75 (m, 3H), 0.71 (t, J=8.0 Hz, 3H).

Example 4 Synthesis of Compound 20

Compound 20: Using an analogous procedure to the procedure in FIG. 5, compound 20 was prepared from compound 16 and azide 2. MS positive mode: m/z=1008.3 [(M+Na)]⁻ C₄₅H₇₅N₇O₁₇ (985).

¹H NMR (400 MHz, D₂O) δ 8.30 (s, 1H), 4.89 (d, J=4.0 Hz, 1H), 4.77 (q, J=8.0 Hz, 1H), 4.46-4.32 (m, 3H), 4.24 (q, J=8.0 Hz, 1H), 4.15 (q, J=8.0 Hz, 1H), 4.05-3.98 (m, 1H), 3.94 (q, J=8.0 Hz, 2H), 3.89-3.73 (m, 1H), 3.75 (dd, J=12.0, 4.0 Hz, 1H), 3.72-3.70 (m, 1H), 3.69-3.64 (m, 2H), 3.63-3.55 (m, 3H), 3.54-3.47 (m, 1H), 3.46-3.31 (m, 9H), 3.30-3.21 (m, 1H), 3.17 (br t, J=12.0 Hz, 1H), 2.30-2.11 (m, 3H), 2.07-1.97 (m, 1H), 1.91 (s, 3H), 1.78-1.31 (m, 9H), 1.30-1.19 (m, 3H), 1.18-1.00 (m, J=8.0 Hz, 7H), 0.99-0.70 (m, 4H), 0.66 (t, J=8.0 Hz, 3H).

Example 5 Synthesis of Compound 21

Compound 21: Using an analogous procedure to the procedure in FIG. 5, compound 20 was prepared from compound 16 and azide 3-azidopropanol. MS positive mode: m/z=948.3 [(M+Na)]⁻ C₄₃H₇₁N₇O₁₅ (925).

¹H NMR (400 MHz, D₂O) δ 8.31 (s, 1H), 7.82 (s, 1H), 4.89 (d, J=4.0 Hz, 1H), 4.78 (q, J=8.0 Hz, 1H), 4.47 (t, J=8.0 Hz, 2H), 4.39 (t, J=8.0 Hz, 2H), 4.24 (q, J=8.0 Hz, 1H), 4.15 (q, J=8.0 Hz, 1H), 4.05-3.99 (m, 1H), 3.94 (q, J=8.0 Hz, 2H), 3.89-3.79 (m, 1H), 3.79-3.64 (m, 6H), 3.63-3.54 (m, 3H), 3.53-3.33 (m, 8H), 3.31-3.22 (m, 1H), 3.17 (br t, J=12.0 Hz, 1H).

Example 6 Prophetic Synthesis of Compound 55

Compound 51: Compound 51 can be prepared in an analogous fashion to FIG. 3 by substituting 2-(proparglyoxy)ethanamine for ethylenediamine.

Compound 55: Compound 55 can be prepared by the same procedure seen in FIG. 5 by substituting compound 51 for compound 12.

Example 7 Prophetic Synthesis of Compound 56

Compound 56: Compound 56 can be prepared by the same procedure seen in FIG. 5 substituting compound 51 for compound 12 and substituting pentyn-5-ol for compound 3.

Example 8 Prophetic Synthesis of Compound 57

Compound 53: Compound 53 can be prepared in an analogous fashion to FIG. 4 by substituting 6-(2-propyn-1-yloxy)-3-pyridinecarboxylic acid for propiolic acid.

Compound 57: Compound 57 can be prepared by the same procedure seen in FIG. 5 substituting compound 53 for compound 12 and substituting compound 4 for compound 3.

Example 9 Prophetic Synthesis of Compound 58

Compound 54: Compound 54 can be prepared in an analogous fashion to FIG. 4 by substituting 5-ethynyl-3-pyridinecarboxylic acid for propiolic acid.

Compound 58: Compound 58 can be prepared by the same procedure as seen in FIG. 5 by substituting compound 54 for compound 12 and substituting 3-azidopropanol for compound 3.

Example 10 Synthesis of Compound 24

Compound 6: Prepared according to Bioorg. Med. Chem. Lett. 1995, 5, 2321-2324 starting with D-threonolactone.

Compound 7: Compound 6 (500 mg, 1 mmol) was dissolved in 9 mL acetonitrile. Potassium hydroxide (1 mL of a 2M solution) was added and the reaction mixture was stirred at 50° C. for 12 hours. The reaction mixture was partitioned between dichloromethane and water. The phases were separated and the aqueous phase was extracted 3 times with dichloromethane. The aqueous phase was acidified with 1N HCl until pH˜1 and extracted 3 times with dichloromethane. The combined dichloromethane extracts from after acidification of the aqueous phase were concentrated in vacuo to give compound 7 as a yellow oil (406 mg). LCMS (C-18; 5-95 H₂O/MeCN): UV (peak at 4.973 min), positive mode: m/z=407 [M+H]⁺; negative mode: m/z=405 [M−H]⁻ C₂₅H₂₆O₅ (406).

Compound 22: HATU (52 mg, 0.14 mmol) and DIPEA (28 μL, 0.16 mmol) were added to a solution of compound 7 (54 mg, 0.13 mmol) in DMF (5 mL) cooled to 0° C. The mixture was stirred at 0° C. for 5 min. A solution of compound 11 (78 mg, 0.106 mmol) in water (2 mL) was then added and the yellow solution was stirred at 0° C.→r.t. for 12 h. The volatiles were evaporated and co-evaporated with toluene (2×). The resulting yellow residue was dried under high vacuum for 3 h. To a solution of the latter residue in pyridine (4 mL) was added acetic anhydride (2 mL) and the solution was stirred at r.t. for 12 h. The volatiles were evaporated and co-evaporated with toluene (2×). The resulting yellow material was purified by column chromatography (DCM/MeOH 0→5%) to give compound 22 as a yellow solid (141 mg). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 5.526 min); UV (peak at 5.432 min).

Compound 23: A suspension of compound 22 (101 mg, 0.08 mmol) and Pd/C 10% (5 mg) in MeOH (10 mL) was degassed (3×) prior to the addition of hydrogen using a balloon. The mixture was stirred at r.t. for 12 h. LCMS analysis shows reaction not complete. The reaction was stirred for further 12 h. LCMS analysis shows reaction gone to completion. The catalyst was filtered off through a short pad of celite and the filtrate concentrated under vacuum to give compound 23 as a white glass (83 mg). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 4.164 min).

Compound 24: Sulfur trioxide pyridine complex was purified as described in the procedure for compound 4. A solution of the purified sulfating reagent (132 mg, 0.83 mmol) and compound 23 (83 mg, 0.08 mmol) in pyridine (3 mL) was stirred at 67° C. for 1 h. The reaction mixture was concentrated under vacuum. The resulting yellow solid was dissolved in water and cooled to 0° C. A 1N solution of NaOH was then added slowly until pH˜10 and the latter was freeze dried. The resulting white solid was then dissolved in MeOH, cooled to −15° C. and treated with NaOMe 25% solution in MeOH (0.5 mL). The mixture was stirred between −15° C. and −10° C. for 30 min. The reaction mixture was neutralised by adding HCl 1N until pH˜7. The volatiles were evaporated and the resulting yellow residue was purified by Gel Permeation (water as eluent). The collected fractions were lyophilised to give compound 24 as an off-white solid (50 mg, 61% over 2 steps). MS negative mode: m/z=544.4 [(M-4Na)/2]⁻ C₃₈H₆₁N₃Na₄O₂₇S₃ (1180).

¹H NMR (300 MHz, D₂O) δ 4.98 (d, J=4.0 Hz, 1H), 4.91-4.81 (m, 2H), 4.47 (d, J=8.2 Hz, 1H), 4.28 (dd, J=10.7, 5.0 Hz, 1H), 4.18 (dd, J=10.7, 7.3 Hz, 1H), 3.98 (t, J=9.4 Hz, 1H), 3.88 (s, 1H), 3.84 (dd, J=10.7, 3.1 Hz, 1H), 3.77-3.73 (m, 1H), 3.72-3.69 (m, 1H), 3.67 (d, J=5.6 Hz, 2H, 3.54 (t, J=5.8 Hz, 1H), 3.46 (d, J=8.6 Hz, 1H), 3.42-3.32 (m, 2H), 3.27 (t, J=9.6 Hz, 2H), 2.31 (t, J=12.4 Hz, 1H), 2.18 (d, J=11.6 Hz, 1H), 2.02 (s, 3H), 1.87-1.73 (m, 3H), 1.72-1.62 (m, 2H), 1.62-1.40 (m, 5H), 1.41-1.22 (m, 2H), 1.18 (d, J=6.5 Hz, 3H), 0.96-0.85 (m, 1H), 0.81 (t, J=7.2 Hz, 3H).

Example 11 Synthesis of Compound 25

Compound 8: Prepared in an analogous fashion to compound 7 using L-erythronolactone as the starting material. LCMS (C-18; 5-95 H₂O/MeCN): ELSD (5.08 min), UV (peak at 4.958 min), positive mode: m/z=407 [M+H]⁺; negative mode: m/z=405 [M−H]⁻ C₂₅H₂₆O₅ (406).

Compound 25: Compound 25 was prepared using an analogous sequence to compound 24, substituting compound 8 for compound 7. MS negative mode: m/z=544.8 [(M-4Na)/2]⁻ C₃₈H₆₁N₃Na₄O₂₇S₃ (1180).

¹H NMR (300 MHz, D₂O) δ 4.97 (d, J=4.0 Hz, 1H), 4.95 (d, J=4.0 Hz, 1H), 4.88-4.78 (m, 2H), 4.47 (d, J=8.6 Hz, 1H), 4.26 (dd, J=10.7, 5.9 Hz, 1H), 4.18 (dd, J=10.6, 5.2 Hz, 1H), 3.97 (dd, J=10.4, 8.5 Hz, 1H), 3.87-3.77 (m, 3H), 3.77-3.71 (m, 2H), 3.71-3.61 (m, 3H), 3.53 (t, J=5.9 Hz, 1H), 3.47-3.36 (m, 1H), 3.36-3.20 (m, 4H), 2.29 (t, J=12.4 Hz, 1H), 2.15 (d, J=12.4 Hz, 1H), 2.01 (s, 3H), 1.88-1.47 (m, 6H), 1.43 (dt, J=8.6, 4.2 Hz, 1H), 1.38-1.21 (m, 3H), 1.18 (d, J=6.6 Hz, 3H), 1.15-1.05 (m, 2H), 0.97-0.84 (m, 1H), 0.80 (t, J=7.3 Hz, 3H).

Example 12 Prophetic Synthesis of Compound 59

Compound 48: Compound 48 can be prepared in an analogous fashion to compound 11, substituting 2, 2′-oxybis(ethylamine) for ethylenediamine.

Compound 59: Compound 59 can be prepared using an analogous sequence to compound 24, substituting compound 48 for compound 11.

Example 13 Prophetic Synthesis of Compound 60

Compound 49: Compound 49 can be prepared in an analogous fashion to compound 11, substituting butylenediamine for ethylenediamine.

Compound 60: Compound 60 can be prepared using an analogous sequence to compound 24, substituting compound 49 for compound 11.

Example 14 Synthesis of Compound 30

Compound 27: Crotonic acid (1.15 g, 13.3 mmol) was dissolved in 30 mL of water. Sodium sulfite (2.03 g, 16 mmol) was added and the reaction mixture was stirred for 1 hour at 80° C. The reaction mixture was cooled on an ice bath and adjusted to pH=1 with concentrated HCl. The solvent was removed in vacuo to afford crude compound 27 which was used without further purification. MS negative mode: m/z=166.9 [M−H]⁻ C₄H₈O₅S (168.17).

¹H NMR (400 MHz, D₂O) δ 3.35 (s, 1H), 2.90 (dd, J=15.9, 5.3 Hz, 1H), 2.47 (dd, J=15.9, 8.7 Hz, 1H), 1.33 (d, J=6.9 Hz, 3H).

Compound 30: To a solution of compound 27 (62 mg, 0.37 mmol) in water was added DIPEA (68 μL, 0.39 mmol), EDCI (75 mg, 0.39 mmol), and NHS (64 mg, 0.56 mmol). The reaction mixture was stirred at room temperature for 2 hours. Compound 11 (184 mg, 0.25 mmol) was added and the reaction mixture was stirred at room temperature overnight. The reaction mixture was directly transferred to a 10 g C-18 cartridge and eluted with water followed by 3/1 mater/methanol. Product containing fractions were combined, concentrated, and lyophilized to afford 20 mg of compound 30 as a white solid. MS negative mode: m/z=903.7 [M−Na]⁻, 881.7 [M-2Na]⁻, 440.5 [(M-2Na)/2]⁻ C₃₈H₆₃N₃Na₂O₁₈S (927.36).

¹H NMR (400 MHz, Deuterium Oxide) δ 5.04 (d, J=4.0 Hz, 1H), 4.91 (q, J=6.4 Hz, 1H), 4.55 (d, J=8.8 Hz, 1H), 3.95 (dd, J=9.4, 3.6 Hz, 1H), 3.89 (dd, J=14.6, 3.3 Hz, 2H), 3.80 (dd, J=10.8, 3.7 Hz, 2H), 3.76-3.70 (m, 2H), 3.55 (t, J=6.0 Hz, 1H), 3.50 (t, J=6.2 Hz, 2H), 3.40-3.28 (m, 3H), 3.13 (t, J=6.1 Hz, 2H), 2.44-2.31 (m, 1H), 2.23-2.11 (m, 1H), 2.06 (d, J=4.4 Hz, 3H), 1.99-1.81 (m, 2H), 1.73 (q, J=13.9 Hz, 1H), 1.66-1.46 (m, 2H), 1.45-1.31 (m, 1H), 1.29 (d, J=6.9 Hz, 1H), 1.24 (d, J=6.4 Hz, 3H), 1.23-1.12 (m, 3H), 1.01-0.90 (m, 2H), 0.86 (t, J=7.3 Hz, 3H).

Example 15 Synthesis of Compound 31

Compound 28: Compound 28 was prepared in analogous fashion to compound 27, starting with 4-bromocrotonic acid and using 2.4 equivalents of sodium sulfite. MS negative mode: m/z=247.1 [M−H]⁻ C₄H₈O₈S₂ (247.97).

¹H NMR (400 MHz, Deuterium Oxide) δ 3.85-3.74 (m, 1H), 3.45 (dd, J=14.2, 1.9 Hz, 1H), 3.03 (dd, J=14.3, 10.9 Hz, 1H), 2.96 (dd, J=16.6, 2.9 Hz, 1H), 2.63 (dd, J=16.6, 9.4 Hz, 1H).

Compound 31: Compound 31 was prepared in an analogous fashion to compound 30, from compound 11 and compound 28. MS negative mode: m/z=1007.5 [M-Na]⁻, 986.5 [M-2Na]⁻, 962.5 [M-3Na]⁻ C₃₈H₆₂N₃Na₃O₂₁S₂ (1029.30).

¹H NMR (400 MHz, Deuterium Oxide) δ 5.03 (d, J=4.1 Hz, 1H), 4.92 (d, J=6.8 Hz, 1H), 4.56 (t, J=8.1 Hz, 1H), 4.03 (t, J=9.6 Hz, 1H), 3.90 (dd, J=10.6, 3.3 Hz, 1H), 3.86 (d, J=3.3 Hz, 1H), 3.80 (dd, J=8.9, 3.5 Hz, 1H), 3.77 (d, J=4.0 Hz, 1H), 3.73 (d, J=6.0 Hz, 2H), 3.60 (t, J=5.9 Hz, 1H), 3.48 (ddd, J=14.2, 5.3, 2.0 Hz, 2H), 3.43-3.36 (m, 1H), 3.33 (t, J=9.8 Hz, 2H), 3.30-3.21 (m, 1H), 3.11-3.05 (m, 1H), 3.05-2.99 (m, 1H), 2.36 (t, J=12.5 Hz, 1H), 2.23 (d, J=11.8 Hz, 1H), 2.07 (s, 3H), 1.89 (d, J=12.7 Hz, 1H), 1.79 (m, 1H), 1.67 (d, J=20.5 Hz, 1H), 1.58 (d, J=16.5 Hz, 1H), 1.53-1.42 (m, 1H), 1.34 (q, J=11.1, 10.2 Hz, 1H), 1.24 (d, J=6.5 Hz, 3H), 1.17 (q, J=12.4, 10.1 Hz, 2H), 1.02-0.91 (m, 1H), 0.87 (t, J=7.5 Hz, 3H).

Example 16 Synthesis of Compound 32

Compound 29: Compound 29 was prepared in analogous fashion to compound 27, starting with 4-bromocrotonic acid and using 1.05 equivalents of sodium sulfite. MS negative mode: m/z=901.7 [M−Na]⁻, 879.7 [M−2Na]⁻ , 439.5 [(M−2Na)/2]⁻² C₃₈H₆₁N₃Na₂O₁₈S (925.35).

¹H NMR (400 MHz, Deuterium Oxide) δ 6.73 (dt, J=15.4, 7.6 Hz, 1H), 6.11 (dd, J=15.6, 1.4 Hz, 1H), 3.81 (d, J=7.4 Hz, 2H).

Compound 32: Compound 32 was prepared in an analogous fashion to compound 30, from compound 11 and compound 29. MS negative mode: m/z=896.3 [M−2Na]⁻ C₃₉H₆₅₁N₃Na₂O₁₈S (941.38).

¹H NMR (400 MHz, Deuterium Oxide) δ 6.69 (dt, J=15.3, 7.6 Hz, 1H), 6.19 (d, J=15.4 Hz, 1H), 5.02 (d, J=4.0 Hz, 1H), 4.90 (q, J=8.5, 7.5 Hz, 1H), 4.52 (d, J=8.6 Hz, 1H), 3.99 (t, J=9.6 Hz, 1H), 3.91-3.81 (m, 4H), 3.81-3.75 (m, 3H), 3.72 (d, J=5.9 Hz, 3H), 3.56 (t, J=5.9 Hz, 1H), 3.48-3.32 (m, 6H), 3.29 (q, J=7.1, 5.8 Hz, 4H), 3.13 (dq, J=22.3, 7.5 Hz, 7H), 2.94 (s, 5H), 2.48 (s, 2H), 2.31 (t, J=12.7 Hz, 1H), 2.13 (d, J=5.4 Hz, 1H), 2.05 (s, 3H), 1.94 (p, J=6.0 Hz, 3H), 1.84 (d, J=11.5 Hz, 1H), 1.78-1.66 (m, 1H), 1.66-1.42 (m, 3H), 1.41-1.28 (m, 1H), 1.23 (d, J=6.4 Hz, 3H), 1.15 (t, J=7.3 Hz, 7H), 1.06 (t, J=7.3 Hz, 1H), 0.93 (q, J=11.3 Hz, 1H), 0.84 (t, J=7.3 Hz, 3H).

Example 17 Synthesis of Compound 34

Compound 34: Compound 34 was prepared in an analogous fashion to compound 30, from compound 11 and the commercially available 5-sulfopentanoic acid. MS negative mode: m/z=896.3 [M−2Na]⁻ C₃₉H₆₅₁N₃Na₂O₁₈S (941.38).

¹H NMR (400 MHz, D₂O) δ 4.93 (d, J=4.0 Hz, 1H), 4.81 (dd, J=8.0 Hz, J=12.0 Hz, 1H), 4.43 (broad d, 1H), 3.95-3.84 (m, 1H), 3.81-3.74 (m, 2H), 3.73-3.68 (m, 2H), 3.67-3.58 (m, 4H), 3.50-3.44 (broad t, 1H), 3.40-3.30 (m, 1H), 2.28-2.11 (m, 5H), 2.85-2.75 (m, 2H), 2.31-2.11 (m, J=8.0 Hz, 3H), 2.10-2.00 (m, 1H), 1.96 (s, 3H), 1.84-1.70 (m, 2H), 1.69-1.34 (m, 11H), 1.33-1.18 (m, 3H), 1.17-0.94 (m, J=8.0 Hz, 8H), 0.93-0.69 (m, J=8.0 Hz, 5H).

Example 18 Synthesis of Compound 33

Compound 33: Compound 32 (14 mg, 0.016 mmol) was dissolved in 1 mL of water. Three drops of saturated NaHCO₃ solution were added followed by Pd/C (4 mg). The reaction mixture was vigorously stirred under a hydrogen atmosphere for 90 minutes. The reaction mixture was filtered through a 0.2 micron PTFE filter. The filtrate was applied to a 5 g C-18 cartridge and eluted with water followed by 3/1 water/methanol. Product containing fractions were concentrated then lyophilized to afford 9 mg of compound 33 as a white solid (64%). MS negative mode: m/z=903.6 [M−Na]⁻, 881.5 [M−2Na]⁻, 440.4 [(M−2Na)/2]⁻² C₃₈H₆₃N₃Na₂O₁₈S (927.36).

¹H NMR (400 MHz, Deuterium Oxide) δ 5.04 (d, J=4.0 Hz, 1H), 4.92 (q, J=6.4 Hz, 1H), 4.55 (d, J=8.6 Hz, 1H), 4.01 (t, J=9.5 Hz, 1H), 3.93-3.85 (m, 3H), 3.85-3.77 (m, 2H), 3.73 (d, J=5.9 Hz, 2H), 3.62-3.55 (m, 1H), 3.50-3.38 (m, 2H), 3.38-3.27 (m, 5H), 2.94-2.86 (m, 2H), 2.46-2.28 (m, 4H), 2.16 (d, J=12.3 Hz, 1H), 2.07 (s, 3H), 2.04-1.96 (m, 2H), 1.86 (d, J=12.8 Hz, 2H), 1.72 (dd, J=38.2, 17.7 Hz, 1H), 1.63-1.44 (m, 3H), 1.44-1.27 (m, 2H), 1.24 (d, J=6.5 Hz, 3H), 1.19-1.12 (m, 11H), 1.01-0.90 (m, 1H), 0.86 (t, J=7.3 Hz, 3H).

Example 19 Synthesis of Compound 35

Compound 35: To a solution of compound 11 (29 mg, 39 μmole) in anhydrous DMSO (0.1 mL) was added a drop of DIPEA and the solution was stirred at room temperature until a homogeneous solution was obtained. A solution of succinic anhydride (4.1 mg, 41.2 μmole, 1.05 eq) in anhydrous DMSO (0.1 mL) was added in one portion and the resulting solution was stirred overnight. The solution was lyophilized to dryness and the crude product was purified by HPLC to give compound 35 (28 mg, 34 μmole, 84%). MS: Calculated (C₃₈H₆₃N₃O₁₇, 833.4), ES-positive (856.3, M+Na), ES-negative (832.3, M-1).

¹H NMR (600 MHz, D₂O) δ 4.92 (d, J=4.0 Hz, 1H), 4.80 (dd, J=8.0 Hz, 1H), 4.42 (d, J=8.0 Hz, 1H), 3.95-3.83 (m, 2H), 3.82-3.73 (m, 2H), 3.72-3.56 (m, 5H), 3.49-3.41 (m, 1H), 3.40-3.31 (m, 1H), 3.30-3.11 (m, 5H), 2.55 (t, J=8.0 Hz, 2H), 2.41 (t, J=8.0 Hz, 2H), 2.26-2.12 (m, 1H), 2.10-1.98 (m, 1H), 1.95 (s, 3H), 1.84-1.68 (m, 2H), 1.67-1.34 (m, 8H), 1.33-1.17 (m, 3H), 1.16-0.92 (m, J=8.0 Hz, 7H), 0.91-0.66 (m, J=8.0 Hz, 5H).

Example 20 Synthesis of Compound 36

Compound 36: Compound 36 was prepared in an analogous fashion to compound 35, from compound 11 and glutaric anhydride. MS: Calculated (C₃₉H₆₅N₃O₁₇, 847.4), ES-positive (870.4, M+Na), ES-negative (846.3, M-1).

¹H NMR (400 MHz, D₂O) δ 4.98 (d, J=4.0 Hz, 1H), 4.86 (dd, J=8.0 Hz, J=12.0 Hz, 1H), 4.46 (broad d, 1H), 4.02 (dd, J=4.0 Hz, J=12.0 Hz, 1H), 3.99-3.91 (m, 1H), 3.89 (m, 1H), 3.83 (dd, J=4.0 Hz, J=8.0 Hz, 1H), 3.77-3.61 (m, 5H), 3.53-3.42 (m, 2H), 3.35-3.18 (m, 5H), 2.34 (t, J=8.0 Hz, 2H), 2.31-2.18 (m, J=8.0 Hz, 3H), 2.13-2.04 (m, 1H), 2.01 (s, 3H), 1.87-1.74 (m, J=4.0 Hz, 4H), 1.73-1.41 (m, 8H), 1.40-1.01 (m, J=8.0 Hz, 10H), 0.96-0.75 (m, J=8.0 Hz, 5H).

Example 21 Prophetic Synthesis of Compound 61

Compound 50: Compound 50 can be prepared in an analogous fashion to compound 11, substituting 1,4-xylylenediamine for ethylenediamine.

Compound 61: Compound 61 can be prepared in an analogous fashion to compound 35, from compound 50 and succinic anhydride.

Example 22 Synthesis of Compound 43

Compound 39: To a solution of compound 38 (445 mg, 1.43 mmol) dissolved in DMF (29 mL) at 0° C. was added HATU (563 mg, 1.14 mmol) and DIPEA (298 μL, 1.71 mmol). The mixture was stirred at 0° C. for 5 min. A solution of compound 11 (840 mg, 1.14 mmol) in water (12 mL) was then added and the mixture was stirred for 12 h allowing to warm to room temperature. The volatiles were evaporated and the crude reaction mixture was co-evaporated 2× with toluene. The resulting yellow residue was dried under high vacuum for 3 h. The residue was dissolved in pyridine (30 mL) at room temperature. Acetic anhydride (15 mL) was added and the solution was stirred at r.t. for 12 h. The volatiles were evaporated and the crude reaction mixture was co-evaporated 2× with toluene. The resulting mixture was purified by column chromatography (DCM/MeOH 0→10%) to give compound 39 (1.197 g, 89%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 4.804 min); UV (peak at 4.693 min).

Compound 40: A suspension of 39 (1.29 g, 1.096 mmol) and Pd/C 10% (50 mg) in MeOH (45 mL) was degassed (3×) prior to the addition of hydrogen using a balloon. The mixture was stirred at r.t. for 12 h. The catalyst was filtered off through a short pad of celite and the filtrate was concentrated under vacuum to give compound 40 as a white solid (1.13 g, 99%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 3.998 min).

Compound 41: To a solution of compound 7 (140 mg, 0.345 mmol) dissolved in DMF (14 mL) and cooled at 0° C. was added HATU (143 mg, 0.38 mmol) and DIPEA (76 μL, 0.435 mmol). The mixture was stirred at 0° C. for 5 min. A solution of compound 40 (300 mg, 0.29 mmol) in DMF (4 mL) was then added and the mixture was stirred overnight allowing to warm to room temperature. The volatiles were evaporated and the residue co-evaporated 2× with toluene. The resulting oil was purified by column chromatography (DCM/MeOH 0→10%) to give compound 41 (317 mg, 76%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 5.352 min); UV (peak at 5.215 min).

Compound 42: A suspension of compound 41 (230 mg, 0.16 mmol) and Pd/C 10% (10 mg) in MeOH (10 mL) was degassed (3×) prior to the addition of hydrogen using a balloon. The mixture was stirred at r.t. for 16 h. Added more Pd/C 10% (10 mg) and stirred for additional 16 h. Reaction was not complete. Added more Pd/C (10 mg) and stirred for another 4 days. The catalyst was filtered off through a short pad of celite and the filtrate concentrated under vacuum. The resulting residue was purified by column chromatography (DCM/MeOH 0-30%) to give compound 42 (123 mg, 67%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 4.055 min).

Compound 43: A solution of sulfur trioxide pyridine (170 mg, 1.067 mmol) and compound 43 (124 mg, 0.106 mmol) in pyridine (4 mL) was stirred at 67° C. for 30 min. The reaction mixture was concentrated under vacuum. The resulting yellow crude was dissolved in water and cooled to 0° C. A 1N solution of NaOH was then added slowly until pH-10 and the mixture stirred overnight allowing to warm to room temperature. The aqueous solution was lyophilized. The resulting white solid was dissolved in MeOH (5 mL), cooled to 0° C. and treated with NaOMe 25% solution in MeOH (0.5 mL). The mixture was stirred between 0° C. and 10° C. for 1 h. The reaction mixture then neutralised by addition of 1N HCl until pH-7. The MeOH was removed in vacuo. Water was added to the residue and the solution was lyophilized. The resulting white solid was purified by C18 ultra SNAP 30 g (3-5% MeOH/H₂O). Product containing fractions were combined and lyophilized to afford compound 43 as an off-white solid (44.5 mg, 18% over 3 steps). LCMS: (C-18; 5-95 H₂O/MeCN): ELSD (peak at 3.039 min), negative mode: m/z=584 [(M-2)/2]⁻ (loss of one sulfate during ionisation), 624 [(M-2)/2]⁻. C₄₅H₇₈N₄O₃₀S₃ (1250) for compound 43 and C₄₅H₇₈N₄O₂₇S₂ (1170) for disulfate.

¹H NMR (300 MHz, Deuterium Oxide) δ 5.01 (d, J=4.0 Hz, 1H), 4.91 (app d, J=6.2 Hz, 2H), 4.51 (d, J=8.6 Hz, 1H), 4.36-4.15 (m, 2H), 3.99 (t, J=9.5 Hz, 1H), 3.92-3.68 (m, 7H), 3.68-3.54 (m, 5H), 3.49-3.37 (m, 3H), 3.31-3.28 (m, 5H), 2.53 (t, J=5.4 Hz, 2H), 2.32 (t, J=12.5 Hz, 1H), 2.16 (app d, J=12.3 Hz, 1H), 2.04 (s, 3H), 1.88-1.67 (m, 2H), 1.65-1.45 (m, 3H), 1.39-1.25 (m, 1H), 1.22 (d, J=6.4 Hz, 3H), 1.18-1.12 (m, 1H), 0.97-0.87 (m, 1H), 0.84 (t, J=7.4 Hz, 4H).

Example 23 Synthesis of Compound 44

Compound 44: Using the sequence of reactions in FIG. 12, compound 44 was prepared substituting compound 8 for compound 7. LCMS: (C-18; 5-95 H₂O/MeCN): ELSD (peak at 3.053 min), negative mode: m/z=584 [(M-2)/2]⁻ (loss of one sulfate during ionisation), 624 [(M-2)/2]⁻. C₄₅H₇₈N₄O₃₀S₃ (1250) for compound 44 and C₄₅H₇₈N₄O₂₇S₂ (1170) for disulfate.

¹H NMR (300 MHz, Deuterium Oxide) δ 5.07 (d, J=4.2 Hz, 2H), 4.96 (q, J=6.5 Hz, 1H), 4.93-4.83 (m, 1H), 4.57 (d, J=8.5 Hz, 1H), 4.31 (qd, J=10.8, 5.4 Hz, 2H), 4.05 (t, J=9.5 Hz, 1H), 3.95-3.88 (m, 2H), 3.87-3.73 (m, 3H), 3.73-3.58 (m, 6H), 3.55-3.43 (m, 3H), 3.36 (s, 4H), 2.58 (t, J=6.1 Hz, 2H), 2.38 (t, J=12.5 Hz, 1H), 2.22 (app d, J=12.3 Hz, 1H), 2.09 (s, 3H), 1.90 (app d, J=12.0 Hz, 1H), 1.85-1.58 (m, 2H), 1.58-1.32 (m, 1H), 1.28 (d, J=6.4 Hz, 3H), 1.24-1.16 (m, 1H), 1.05-0.94 (m, 1H), 0.90 (t, J=7.3 Hz, 3H).

Example 24 Synthesis of Compound 46

Compound 45: To a stirred and cooled (15° C.) solution of propiolic acid (82 μl, 1.19 mmol) in DCM (5 mL) under argon was added oxalyl chloride (100 μl, 1.19 mmol) and DMF (5 μl, cat). The resulting mixture was stirred for 20 min. A solution of compound 40 (180 mg, 0.17 mmol) in MeOH (20 mL) was treated with saturated sodium bicarbonate solution (4 mL) 1 min prior to the addition of 3.5 ml of the propiolyl chloride solution. The resulting mixture was stirred at room temperature for 40 min. The reaction was filtered and partially concentrated under reduced pressure to the water fraction, which, was in-turn freeze-dried. Flash-column chromatography of the residue (0-10% MeOH in DCM) afforded compound 45 as a white solid (110 mg, 60%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 3.38 min), UV (peak at 3.29 min), negative mode: m/z=1149 [M+Na] C₅₃H₈₂N₄O₂₂ (1126).

Compound 46: A stirred solution of compound 45 (110 mg, 0.1 mmol) in MeOH (20 ml) under argon was treated with sodium methoxide until pH=9-10 then stirred for 2 h. HPLC grade water (10 mL) was added and the mixture concentrated by rotary evaporation. The resulting aqueous solution was lyophilized. The residue was dissolved in HPLC grade water (20 ml) and the pH adjusted to 9-10 with 1N NaOH and stirred at room temperature for 2 h. The pH was adjusted to 8 (1M HCl) and the mixture lyophilized. Salts were removed by G10 gel permeation of the residue, (water as eluent).

The residue was combined with azide 3 in water (15 mL) under argon at room temperature. A pre-mixed solution of copper sulfate and THPTA (0.43 mL of 40 mmol aqueous solution) and sodium ascorbate (107 μmol, 0.107 mL of 1M solution) was added and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was lyophilized. The crude reaction mixture was partially purified by P2 gel permeation (water as eluent) to give a yellow solid. This material was further purified by Biotage reverse phase C18 ultra (30 g) (3-5% H₂O/MeOH) chromatography afforded compound 46 (54 mg, 46%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 3.08 min), UV (peak at 2.98 min), negative mode: m/z=671.2 [(M-2)/2]⁻ C₄₉H₈₂N₇O₃₀S₃ (1344).

¹H NMR (300 MHz, Deuterium Oxide) δ 8.47 (s, 1H), 5.01 (d, J=4.0 Hz, 1H), 4.90 (q, J=6.7 Hz, 1H), 4.71 (s, 2H), 4.51 (d, J=8.6 Hz, 1H), 4.05 (s, 6H), 3.98 (t, J=9.4 Hz, 1H), 3.92-3.77 (m, 5H), 3.76-3.63 (m, 8H), 3.57 (dt, J=11.3, 5.5 Hz, 3H), 3.43 (d, J=10.8 Hz, 1H), 3.36-3.19 (m, 6H), 2.45 (t, J=6.0 Hz, 2H), 2.29 (app t, J=12.2 Hz, 1H), 2.15 (app d, J=11.8 Hz, 1H), 2.04 (s, 3H), 1.82 (app d, J=12.3 Hz, 2H), 1.78-1.42 (m, 2H), 1.47-1.25 (m, 1H), 1.22 (d, J=6.5 Hz, 3H), 1.17-1.07 (m, 1H), 0.97-0.88 (m, 1H), 0.82 (t, J=7.3 Hz, 3H).

Example 25 Prophetic Synthesis of Compound 73

Compound 63: A solution of compound 62 (preparation described in WO 2007/028050) and phenyl acetylene (2.4 eq) in MeOH is degassed at room temperature. A solution of CuSO₄/THPTA in distilled water (0.04 M) (0.2 eq) and sodium ascorbate (1.2 eq) is added successively and the resulting solution stirred 12 hrs at room temperature. The solution is concentrated under reduced pressure. The crude product is purified by flash chromatography to give compound 63.

Compound 65: A mixture of compound 63 and compound 64 (preparation described in WO 2013/096926) (1.7 eq) is azeotroped 3× from toluene. The mixture is dissolved in DCM under argon and cooled on an ice bath. To this solution is added boron trifluoride etherate (1.5 eq). The reaction mixture is stirred 12 hours at room temperature. The reaction is quenched by the addition of triethylamine (2 eq). The reaction mixture is transferred to a separatory funnel and washed 1× half saturated sodium bicarbonate solution and 1× water. The organic phase is dried over sodium sulfate, filtered, and concentrated. The residue is purified by flash chromatography to afford compound 65.

Compound 66: Compound 65 is dissolved in methanol at room temperature. A solution of sodium methoxide in methanol (0.1 eq) is added and the reaction mixture stirred overnight at room temperature. The reaction mixture is quenched by the addition of acetic acid. The reaction mixture is diluted with ethyl acetate, transferred to a separatory funnel and washed 2× with water. The organic phase is dried over magnesium sulfate, filtered and concentrated. The residue is separated by flash chromatography to afford compound 66.

Compound 67: To a solution of compound 66 cooled on an ice bath is added DABCO (1.5 eq) followed by monomethyoxytrityl chloride (1.2 eq). The reaction mixture is stirred overnight allowing to warm to room temperature. The reaction mixture is concentrated and the residue is purified by flash chromatography to afford compound 67.

Compound 68: To a solution of compound 67 in methanol is added dibutyltin oxide (1.1 eq). The reaction mixture is refluxed for 3 hours then concentrated. The residue is suspended in DME. To this suspension is added 2-(R)-[[(trifluoromethyl)sulfonyl]oxy]-cyclohexanepropanoic acid benzyl ester (preparation described in Thoma et. al. J. Med. Chem., 1999, 42, 4909) (1.5 eq) followed by cesium fluoride (1.2 eq). The reaction mixture is stirred at room temperature overnight. The reaction mixture is diluted with ethyl acetate, transferred to a separatory funnel, and washed with water. The organic phase is dried over sodium sulfate, filtered and concentrated. The residue is purified by flash chromatography to afford compound 68.

Compound 69: To a degassed solution of compound 68 in anhydrous DCM at 0° C. is added Pd(PPh₃)₄ (0.1 eq), Bu₃SnH (1.1 eq) and acetic anhydride (2.0 eq). The resulting solution is stirred for 12 hrs while the temperature is gradually increased to room temperature. The reaction mixture is diluted with DCM, transferred to a separatory funnel, and washed with water. The organic phase is dried over Na₂SO₄, then filtered and concentrated. The residue is purified by flash chromatography to afford compound 69.

Compound 70: Compound 69 is dissolved in methanol and degassed. To this solution is added Pd(OH)₂/C. The reaction mixture is vigorously stirred under a hydrogen atmosphere for 12 hours. The reaction mixture is filtered through a Celite pad and the cake is washed with MeOH. The combined filtrate is concentrated under reduced pressure. The crude product is washed with hexane and dried under high vacuum to give compound 70.

Compound 71: Compound 70 is dissolved in ethylenediamine and stirred at 70° C. for 8 hours. The solvent is removed in vacuo and the crude material separated by C-18 reverse phase chromatography to afford compound 71.

Compound 72: To a stirred and cooled (15° C.) solution of propiolic acid in DCM under argon is added oxalyl chloride and DMF (cat). The resulting mixture is stirred for 20 min. A solution of compound 72 in MeOH containing saturated sodium bicarbonate solution is added. The resulting mixture is stirred at room temperature for 30 min. The reaction is filtered and concentrated under reduced pressure. Gel permeation of the residue (water as eluent) affords compound 72.

Compound 73: A stirred solution of compound 72 and compound 3 in water under argon is treated with a pre-mixed solution of copper sulfate and THPTA (40 mM aqueous solution) and sodium ascorbate. Upon completion the reaction is lyophilized. Gel permeation of the residue (water as eluent) affords compound 73.

Example 26 E-Selectin Activity—Binding Assay

The inhibition assay to screen for and characterize glycomimetic antagonists of E-selectin is a competitive binding assay, which allows the determination of IC₅₀ values. E-selectin/Ig chimera was immobilized in 96 well microtiter plates by incubation at 37° C. for 2 hours. To reduce nonspecific binding, bovine serum albumin was added to each well and incubated at room temperature for 2 hours. The plate was washed and serial dilutions of the test compounds were added to the wells in the presence of conjugates of biotinylated, sLe^(a) polyacrylamide with streptavidin/horseradish peroxidase and incubated for 2 hours at room temperature.

To determine the amount of sLe^(a) bound to immobilized E-selectin after washing, the peroxidase substrate, 3,3′,5,5′ tetramethylbenzidine (TMB) was added. After 3 minutes, the enzyme reaction was stopped by the addition of H₃PO₄, and the absorbance of light at a wavelength of 450 nm was determined. The concentration of test compound required to inhibit binding by 50% was determined and reported as the IC₅₀ value for each glycomimetic E-selectin antagonist as shown in the table below. IC₅₀ values for exemplary compounds disclosed herein are provided in the following table.

E-Selectin Antagonist Activity Compound IC₅₀ (nM) 17 150 18 170 24 190 25 660 19 580 20 240 21 520 30 2777 31 2356 33 2115 34 340 36 510 35 720 43 106 44 136 46 192 

What is claimed is:
 1. At least one compound chosen from glycomimetic E-selectin antagonists of Formula (I):

prodrugs of Formula (I), and pharmaceutically acceptable salts of any of the foregoing, wherein R¹ is chosen from H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₁₋₈ haloalkyl, C₂₋₈ haloalkenyl, C₂₋₈ haloalkynyl,

groups, wherein n is chosen from integers ranging from 0 to 2, R⁷ is chosen from H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₄₋₁₆ cycloalkylalkyl, and C(═O)R⁸ groups, and each R⁸ is independently chosen from H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₄₋₁₆ cycloalkylalkyl, C₆₋₁₈ aryl, and C₁₋₁₃ heteroaryl groups; R² is chosen from C₁₋₁₂ alkyl groups substituted with at least one substituent chosen from —OSO₃Q groups, wherein each Q is independently chosen from H and pharmaceutically acceptable cations; R³ is chosen from —OH, —OY¹, halo, —NH₂, —NHY¹, —NY¹Y², —OC(═O)Y¹, —NHC(═O)Y¹, and —NHC(═O)NHY¹ groups, wherein Y¹ and Y², which may be the same or different, are independently chosen from C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₄₋₁₆ cycloalkylalkyl, C₂₋₁₂ heterocyclyl, C₆₋₁₈ aryl, and C₁₋₁₃ heteroaryl groups, wherein Y¹ and Y² may join together along with the nitrogen atom to which they are attached to form a ring; R⁴ is chosen from —CN, —CH₂CN, and —C(═O)Y³ groups, wherein Y³ is chosen from C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, —OZ¹, —NHOH, —NHOCH₃, —NHCN, and —NZ¹Z² groups, wherein Z¹ and Z², which may be identical or different, are independently chosen from H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₁₋₁₂ haloalkyl, C₂₋₁₂ haloalkenyl, C₂₋₁₂ haloalkynyl, and C₇₋₁₂ arylalkyl groups, wherein Z¹ and Z² may join together along with the nitrogen atom to which they are attached to form a ring; R⁵ is chosen from H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₁₋₁₂ haloalkyl, C₂₋₁₂ haloalkenyl, C₂₋₁₂ haloalkynyl, C₄₋₁₆ cycloalkylalkyl, and C₆₋₁₈ aryl groups; R⁶ is chosen from —CN, C₁₋₁₂ alkyl, and C₁₋₁₂ haloalkyl groups; L is chosen from a bond, C₆₋₁₈ aryl, and C₁₋₁₂ heteroaryl groups; and M is chosen from linker groups.
 2. The at least one compound according to claim 1, wherein R¹ is chosen from C₁₋₄ alkyl,


3. The at least one compound according to claim 2, wherein R¹ is chosen from methyl, ethyl,


4. The at least one compound according to claim 1, wherein R³ is chosen from —OH, —OY¹, —OC(═O)Y¹, and —NHC(═O)Y¹ groups, wherein Y¹ is chosen from C₁₋₁₂ alkyl, C₄₋₁₆ cycloalkylalkyl, C₂₋₁₂ heterocyclyl, C₆₋₁₈ aryl, and C₁₋₁₃ heteroaryl groups.
 5. The at least one compound according to claim 4, wherein R³ is chosen from


6. The at least one compound according to claim 5, wherein R³ is chosen from


7. The at least one compound according to claim 1, wherein R⁴ is chosen from —C(═O)Y³ groups, wherein Y³ is chosen from —OZ¹ and —NZ¹Z² groups.
 8. The at least one compound according to claim 7, wherein Z¹ and Z², which may be identical or different, are independently chosen from H, C₁₋₁₂ alkyl, C₁₋₁₂ haloalkyl, and C₇₋₁₂ arylalkyl groups.
 9. The at least one compound according to claim 7, wherein R⁴ is chosen from


10. The at least one compound according to claim 7, wherein R⁴ is chosen from


11. The at least one compound according to claim 1, wherein R⁵ is chosen from C₁₋₁₂ alkyl and C₄₋₁₆ cycloalkylalkyl groups.
 12. The at least one compound according to claim 11, wherein R⁵ is chosen from propyl and cyclohexylmethyl.
 13. The at least one compound according to claim 1, wherein R⁶ is chosen from CN, CF₃, and methyl.
 14. The at least one compound according to claim 13, wherein R⁶ is methyl.
 15. The at least one compound according to claim 1, wherein L is a bond.
 16. The at least one compound according to claim 1, wherein L is chosen from C₆₋₁₈ aryl groups.
 17. The at least one compound according to claim 16, wherein L is:


18. The at least one compound according to claim 1, wherein L is chosen from C₁₋₁₂ heteroaryl groups.
 19. The at least one compound according to claim 18, wherein L is:


20. The at least one compound according to claim 1, wherein the linker group is chosen from:


21. The at least one compound according to claim 1, wherein R² is chosen from C₁₋₁₂ alkyl groups substituted with at least two substituents chosen from —OSO₃Q groups.
 22. The at least one compound according to claim 1, wherein R² is chosen from C₁₋₁₂ alkyl groups substituted with at least three substituents chosen from —OSO₃Q groups.
 23. The at least one compound according to claim 1, wherein R² is chosen from C₁₋₅ alkyl groups substituted with one, two, or three substituents independently chosen from —OSO₃Q groups.
 24. The at least one compound according to claim 1, wherein R² is chosen from C₁₋₈ alkyl groups substituted with one, two, or three substituents independently chosen from —OSO₃Q groups.
 25. The at least one compound according to claim 1, wherein the at least one compound is chosen from

and pharmaceutically acceptable salts of any of the foregoing.
 26. The at least one compound according to claim 1, wherein the at least one compound is chosen from

and pharmaceutically acceptable salts thereof.
 27. The at least one compound according to claim 1, wherein the at least one compound is chosen from


28. The at least one compound according to claim 1, wherein the at least one compound is


29. The at least one compound according to claim 1, wherein each Q is independently chosen from pharmaceutically acceptable cations.
 30. The at least one compound according to claim 1, wherein each Q is a sodium cation.
 31. A composition comprising at least one compound according to claim 1 and at least one additional pharmaceutically acceptable ingredient.
 32. A method for decreasing the likelihood of the occurrence of: or abatement, lessening, decreasing occurrence, or alleviation of symptoms of: or delay or slowing of the progression of: or prolonging the survival of a subject afflicted with: or partial remission of: or ameliorating at least one disease, disorder, and/or condition where inhibition of E-selectin mediated functions is useful, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 33. A method for decreasing the likelihood of the occurrence of: or abatement, lessening, decreasing occurrence, or alleviation of symptoms of: or delay or slowing of the progression of: or prolonging the survival of a subject afflicted with: or partial remission of: or ameliorating at least one inflammatory disease, disorder, and/or condition, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 34. A method for decreasing the likelihood of the occurrence of: or abatement, lessening, decreasing occurrence, or alleviation of symptoms of: or delay or slowing of the progression of: or prolonging the survival of a subject afflicted with; or partial remission of: or ameliorating cancer, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 35. The method according to claim 34, wherein the cancer is chosen from solid tumor cancers.
 36. The method according to claim 34, wherein the cancer is chosen from bone cancers, colorectal cancers, and pancreatic cancers.
 37. The method according to claim 34, wherein the cancer is chosen from liquid tumor cancers.
 38. The method according to claim 34, wherein the cancer is chosen from acute myelogenous leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, and multiple myeloma.
 39. A method for decreasing the likelihood of the occurrence of: or abatement, lessening, decreasing occurrence, or alleviation of symptoms of: or delay or slowing of the progression of: or prolonging the survival of a subject afflicted with: or partial remission of: or ameliorating cancer, the method comprising administering to a subject in need thereof (a) an effective amount of at least one compound of claim 1 and (b) at least one of therapy chosen from (i) chemotherapy and (ii) radiotherapy.
 40. A method for decreasing the likelihood of the occurrence of: or abatement, lessening, decreasing occurrence, or alleviation of symptoms of: or delay or slowing of the progression of: or prolonging the survival of a subject afflicted with: or partial remission of: or ameliorating metastasis of cancer cells, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 41. A method for inhibiting infiltration of cancer cells into the liver, lymph nodes, lung, bone, and/or bone marrow, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 42. A method for enhancing hematopoietic stem cell survival, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 43. The method according to claim 42, wherein the subject has cancer and has received or will receive chemotherapy and/or radiotherapy.
 44. A method for mobilizing cells from the bone marrow, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 45. The method according to claim 44, wherein the cells are chosen from hematopoietic cells and tumor cells.
 46. A method for decreasing the likelihood of the occurrence of: or abatement, lessening, decreasing occurrence, or alleviation of symptoms of: or delay or slowing of the progression of: or prolonging the survival of a subject afflicted with: or partial remission of: or ameliorating mucositis, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 47. The method according to claim 46, wherein the mucositis is chosen from oral mucositis, esophageal mucositis, and gastrointestinal mucositis.
 48. The method according to claim 46, wherein the subject is afflicted with head and neck, breast, lung, ovarian, prostate, lymphatic, leukemic, and/or gastrointestinal cancer.
 49. A method for decreasing the likelihood of the occurrence of: or abatement, lessening, decreasing occurrence, or alleviation of symptoms of: or delay or slowing of the progression of: or prolonging the survival of a subject afflicted with: or partial remission of: or ameliorating thrombosis, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 50. A method for decreasing the likelihood of the occurrence of: or abatement, lessening, decreasing occurrence, or alleviation of symptoms of: or delay or slowing of the progression of: or prolonging the survival of a subject afflicted with: or partial remission of: or ameliorating at least one cardiovascular disease or complications associated therewith, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 51. The method according to claim 50, wherein the at least one cardiovascular disease is chosen from atherosclerosis and myocardial infarction.
 52. A method of inhibiting rejection of a transplanted tissue in a subject, wherein said subject is a recipient of the transplanted tissue, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 53. A method for decreasing the likelihood of the occurrence of: or abatement, lessening, decreasing occurrence, or alleviation of symptoms of: or delay or slowing of the progression of: or prolonging the survival of a subject afflicted with: or partial remission of: or ameliorating graft versus host disease or complications associated therewith, the method comprising administering to a subject in need thereof an effective amount of at least one compound of claim
 1. 